粪便标本基因标记物mRNA表达检测及其对结直肠癌诊断的研究
摘要
结直肠癌(CRC)是最常见的恶性肿瘤之一,也是导致肿瘤病人死亡的重要疾病,其发病率在西方国家占恶性疾病的第二位,在非吸烟者中,结直肠癌是最为严重的致命性癌性疾病之一。在美国,结直肠癌患者占所有癌疾病患者的11%,2001年统计,约有130200新发病例,死亡人数约48100,在日本,每年有新发病例85000人,病死人数约35600人。如果病人没有转移,绝大多数病人可以成功治愈,因此,早期诊断和早期治疗是十分重要的。为了减少结直肠癌的死亡率,寻求早期诊断结直肠癌的普查手段是十分必要的。
目前为止,粪便潜血试验(FOBT)是应用广泛的结直肠癌普查方法。然而最近三项大规模的调查研究显示,以结直肠镜检查为标准,FOBT可作为筛查结直肠癌的无创性普查方法,减少了癌性相关病死率,但其敏感性较差,仅为15%-35%,提示大量的结直肠肿瘤病人可以被漏诊,另外,饮食成分中过氧化物酶的存在也易导致假阳性,试验前必须限定饮食。随着对结直肠癌的分子发生机制的深入理解,粪便中肿瘤源性DNA突发的检测作为结直肠癌无创性筛查手段已成为目前研究热点之一,由于基因的变化直接与肿瘤的发生、发展相关,其明显优越于肿瘤的间接标记物检测,如FOBT,但由于结直肠癌的克隆的异质性,使粪便DNA突变检查的敏感性较差是其不利因素。
几项有关粪便基因mRNA检测技术在结直肠癌诊断中的研究报告表明,采用RT-PCR技术检测结直肠癌病人粪便CD44基因的异常表达对CRC诊断率可达68%。日本学者研究表明,COX-2mRNA在结直肠癌患者粪便中表达率较高,但关于CK20能否在CRC粪便中表达,CD44、COX-2、CK20在病灶及粪便中表达是否具有同源性,联合检测是否能够提高CRC的阳性诊断率,以及三者联合检测能否用于癌性肠梗阻的诊断有待于研究。
研究目的：本实验的目的是通过对COX-2, CK20, CD44v6mRNA在结直肠原发灶及病人粪便中的表达,证实粪便中COX-2, CK20, CD44v6mRNA表达来源于原发灶肿瘤细胞的脱落所致,为临床结直肠癌筛查选择合适的多基因标记物联合检测指标,通过多基因mRNA表达检测癌性肠梗阻病人粪便,观察三种标记物检测定性诊断癌性肠梗阻的价值。
研究方法及结果：我们将实验分成如下步骤：
1.COX-2, CK20, CD44v6mRNA在结直肠癌原发灶及粪便中的表达。对38例行结直肠癌根治术的病人,原发灶及术前收集的粪便标本采用RT-PCR技术对COX-2, CK20, CD44v6mRNA表达进行检查,结果发现,COX-2mRNA在原发灶及粪便中同时阳性表达率为44.7%(17/38),同时阴性表达率为39.5%(15/38),同源性可达84.2%(32/38)；CK20有44.7%(17/38)同时阳性表达,36.8%(14/38)同时无表达,同源性为81.5%(31/38)；CD44v6 mRNA有50%(19/38)同时阳性表达,44.7%(17/38)同时无表达,同源性为94.7%(36/38),同时性表达差异具有显著性,p值均小于0.001,说明粪便组织中COX-2, CK20及CD44v6mRNA表达来源于原发灶脱落的肿瘤细胞。
对38例行结直肠癌根治术病人及22例健康志愿者粪便,采用RT-PCR技术对COX-2, CK20, CD44v6mRNA表达进行检测,结果发现,COX-2mRNA在大肠癌组及健康对照组中阳性率分别为47.4%(18/38),13.6%(3/22),二者相比,差异具有显著性,p=0.0083; CK20mRNA在大肠癌组及健康对照组中阳性率分别为55.3%(21/38),18.2%(4/22),二者相比,差异具有显著性,p=0.0050; CD44v6mRNA在大肠癌组中阳性率为50.0%(19/38),在健康对照组中为9%(2/22),二者相比,差异具有显著性,p=0.0014。提示粪便中COX-2, CK20, CD44v6mRNA表达与大肠癌密切相关,进一步研究发现,三者联合检测可明显提高其阳性检出率,阳性率为86.8%(33/38),联合检测的Se,Sp,PPV,NPV及Ac分别为81.6%,81.8%,88.6%,72%及81.7%。
2.粪便基因标记物COX-2, CK20, CD44v6mRNA联合检测定性诊断肠梗阻中的应用。
对27例通过X线或腹部CT证实为肠梗阻的病人,检测粪便基因标记物mRNA表达,同时手术证实20例为癌性肠梗阻,7例为非癌性肠梗阻,20例癌性肠梗阻病人中有14例伴发肝转移,对癌性肠梗阻的诊断率为90%(18/20),其Se, Sp, PPV, NPV及Ac分别为94.7%,85.7%,90%,85.7%及88.9%。
结论：根据以上结果得出如下结论：
1.粪便COX-2, CK20, CD44v6mRNA表达与大肠癌原发灶中的表达具有同源性,说明粪便中COX-2, CK20, CD44v6mRNA来源于原发灶肿瘤细胞的脱落。
2.检测粪便中COX-2, CK20, CD44v6mRNA表达可用于结直肠癌的诊断,联合检测有助于提高其阳性检出率。
3.通过联合检测粪便COX-2, CK20, CD44v6mRNA表达可作为无法行镜检的癌性肠梗阻病人的无创性诊断手段。
Colorectal cancer (CRC) is one of the most frequent cancers and the leading to death of important disease of tumors. Colorectal cancer is the second common cause of cancer death in the western world, and is the most severe fatal disease among nonsmokers. In the United States, CRC accounts for 11% of all cancers, with an estimated 130200 new cases and 48100 death in 2001, and in Japan, there are 85000 annual cancer registrations and 35600 death caused by CRC. Because most of patients can be treated successfully if metastasis does not occur, it is important to make an early diagnosis and early treatment. In order to reduce the mortality rate of CRC, the development of the screening test by which the cancer can be diagnosed at an early stage is necessary.
To date, the fecal occult blood test(FOBT) has been used widely as the screening test for CRC as a noninvasive way. However, three recent large-scale studies showed that the sensitivity of FOBT was low using total colonoscopy as a reference standard in all subjects. However, the sensitivity of FOBT is quiet low, (15%-35%), which implies that a substantial proportion of colorectal neoplasms may be missed. In addition, the peroxidase activity present in dietary components may lead to falsepositive results, which mandates the need for special dietary restrictions prior to stool collection. With a better understanding of molecular changes associated with CRC development, tumor derived DNA alternations in stool as a noninvasive molecular screening test for colorectal neoplasmas has becoming a "hot spot" of searching. Because genetic alterations are associated directly with the devolopment of neoplasia, they have clear advantages over indirect markers such as FOBT, the disadvantage of DNA-based stool assays is the lack of sensitivity caused by clonal heterogeneity in CRC.
RNA-based stool assays have been reported in several preliminary studies, one of which showed that CD44 variant expression in human feces could be detected in 68% of CRC patients by a combination of reverse-transcription polynerase chain reaction (RT-PCR) and southern hybridization. A group of Japanese investigators has demonstrated the potential of detecting cyclooxygenase-2(COX-2)mRNA, which is frequently overexpressed in the fecal samples patients with colorectal cancer, however, whether CK20mRNA is also detected in the stool samples of patients with colorectal cancers remains unknown, whether COX-2mRNA, CK20mRNA, and CD44v6mRNA expression in the stools samples and primary lesions of CRC have hemogenous expression remains unknown. No investigations have been conducted regarding combined detection of genetic biomarkers RNA-based stool assays could improve positive-diagnosis rate of CRC and diagnose cancer-related intestinal obstruction.
Purpose:The objective of our study is to determine expression of genetic biomarkers RNA-based stool samples originate from the continuously exfoliate tumor cells shed from the primary lesions of colorectal cancers or neoplasmas, by determining expression of COX-2, CK20 and CD44v6mRNA of the primary lesions and fecal samples of CRC, providing suitable combined RNA-based stool assays for noninvasive screening test of colorectal neoplasmas or cancers. The determination of genetic biomarkers RNA-based stool assays and assessed its usefulness as a noninvasive diagnostic tool for determining cancer-related intestinal obstruction.
Method and result:We divide the experiment into several parts:
The part 1:Expression of COX-2, CK20 and CD44v6mRNA in primary colorectal carcinoma lesions and exfoliate tumor cells of fecal samples.
We carry out the expression of COX-2, CK20 and CD44v6mRNA by using RT-PCR in 38 cases of colorectal primary lesions and exfoliate tumor cells of fecal samples, we found that the agreed positive expression rate of COX-2mRNA was 44.7%(17/38) in two groups samples, the agreed negative expression rate of COX-2mRNA was 39.5%(15/38) in them, their homogenous expression rate was 84.2%(32/38); the agreed positive expression rate of CK20mRNA was 44.7%(17/38) in the primary colorectal lesions and stool samples, the agreed negative expression rate of CK20 mRNA was 36.8%(14/38), their homogenous expression rate was 81.5%(31/38); the agreed positive and negative expression rate of CD44v6mRNA in the primary lesions and fecal samples were 50%(19/38),44.7%(17/38), respectively, the homogenous expression were statistically significantly different from hemogenous expression, p<0.001, respectively, it indicated that the expression of genetic biomarkers RNA-based samples originate from the continously exfoliated tumor cells shed from the primary lesions of colorectal cancers.
Fecal samples from 38 patients with colorectal carcinoma before radical resection and from 22 cases of normal healthes severed as controls, we determined the expression of COX-2,CK20 and CD44v6mRNA of fecal exfoliated cells by using RT-PCR assay. We found that the positive expression rate of COX-2mRNA in the group of colorectal cancer and healthy control group were 47.4%(18/38),13.6%(3/22), respectively, the rate was statistically significantly different in two groups, p=0.0083; the postive expression rate of CK20mRNA in the colorectal carcinoma group and healthy control group were 55.3%(21/38),18.2%(4/22), respectively, the rate was statistically significantly different in them, p=0.0050; the postive expression rate of CD44v6mRNA in two groups were 50.0%(19/38),9%(2/22), respectively, the rate was statistially significantly different, p=0.0014, it indicated that the expression of CK20, COX-2 and CD44v6mRNA in fecal samples was closely related to the progresssion of colorectal cancers. The further study found that the combined detection of three genetic biomarkers in fecal samples can improve significantly the positive rate of colorectal cancer, the positive rate was 86.8%(33/38), obviously superior to the positive rate of single genetic biomarkers. p<0.001, the Se, Sp, PPV, NPV and Ac of combined detection of three biomarkers were 81.6%,81.8%,88.6%,72% and 81.7%, respectively.
The part 2:Combined detection of genetic biomarkers COX-2, CK20 and CD44v6mRNA and diagnose to cancer-related intestinal obstruction.
27 patients diagnosed by X-ray or abdominal CT, while 20 patients diagnosed as cancer-related intestinal obstruction by operation. The expression of COX-2, CK20, CD44v6mRNA of fecal samples were assessed by RT-PCR assay. We found that the diagnosis rate of cancer-related intestinal obstruction patients was 90%(18/20), the Se, Sp, PPV, NPV and Ac of combined detection of three genetic biomarkers were 94.7%,85.7%,90%,85.7%and 88.9%, respectively.
Conclusion:From the above date and discussion, we can draw the following conclusion:
1.There is a high homogenous expression rate of COX-2, CK20 and CD44v6mRNA in the primary colorectal cancer lesions and fecal samples, it indicated the expression of genetic biomarkers RNA-based samples originate from the continuously exfoliated tumor cells shed from the primary lesions of colorectal cancer.
2.Combined detection of expression of COX-2, CK20 and CD44v6 mRNA appears to be useful, noninvasive biological markers for diagnosis of colorectal cancer.
3.Combined detection of tumor-derived RNA-based stool assay can be as a noninvasive molecular diagnosis assay for cancer-related intestinal instead of endoscopy.
目前为止,粪便潜血试验(FOBT)是应用广泛的结直肠癌普查方法。然而最近三项大规模的调查研究显示,以结直肠镜检查为标准,FOBT可作为筛查结直肠癌的无创性普查方法,减少了癌性相关病死率,但其敏感性较差,仅为15%-35%,提示大量的结直肠肿瘤病人可以被漏诊,另外,饮食成分中过氧化物酶的存在也易导致假阳性,试验前必须限定饮食。随着对结直肠癌的分子发生机制的深入理解,粪便中肿瘤源性DNA突发的检测作为结直肠癌无创性筛查手段已成为目前研究热点之一,由于基因的变化直接与肿瘤的发生、发展相关,其明显优越于肿瘤的间接标记物检测,如FOBT,但由于结直肠癌的克隆的异质性,使粪便DNA突变检查的敏感性较差是其不利因素。
几项有关粪便基因mRNA检测技术在结直肠癌诊断中的研究报告表明,采用RT-PCR技术检测结直肠癌病人粪便CD44基因的异常表达对CRC诊断率可达68%。日本学者研究表明,COX-2mRNA在结直肠癌患者粪便中表达率较高,但关于CK20能否在CRC粪便中表达,CD44、COX-2、CK20在病灶及粪便中表达是否具有同源性,联合检测是否能够提高CRC的阳性诊断率,以及三者联合检测能否用于癌性肠梗阻的诊断有待于研究。
研究目的：本实验的目的是通过对COX-2, CK20, CD44v6mRNA在结直肠原发灶及病人粪便中的表达,证实粪便中COX-2, CK20, CD44v6mRNA表达来源于原发灶肿瘤细胞的脱落所致,为临床结直肠癌筛查选择合适的多基因标记物联合检测指标,通过多基因mRNA表达检测癌性肠梗阻病人粪便,观察三种标记物检测定性诊断癌性肠梗阻的价值。
研究方法及结果：我们将实验分成如下步骤：
1.COX-2, CK20, CD44v6mRNA在结直肠癌原发灶及粪便中的表达。对38例行结直肠癌根治术的病人,原发灶及术前收集的粪便标本采用RT-PCR技术对COX-2, CK20, CD44v6mRNA表达进行检查,结果发现,COX-2mRNA在原发灶及粪便中同时阳性表达率为44.7%(17/38),同时阴性表达率为39.5%(15/38),同源性可达84.2%(32/38)；CK20有44.7%(17/38)同时阳性表达,36.8%(14/38)同时无表达,同源性为81.5%(31/38)；CD44v6 mRNA有50%(19/38)同时阳性表达,44.7%(17/38)同时无表达,同源性为94.7%(36/38),同时性表达差异具有显著性,p值均小于0.001,说明粪便组织中COX-2, CK20及CD44v6mRNA表达来源于原发灶脱落的肿瘤细胞。
对38例行结直肠癌根治术病人及22例健康志愿者粪便,采用RT-PCR技术对COX-2, CK20, CD44v6mRNA表达进行检测,结果发现,COX-2mRNA在大肠癌组及健康对照组中阳性率分别为47.4%(18/38),13.6%(3/22),二者相比,差异具有显著性,p=0.0083; CK20mRNA在大肠癌组及健康对照组中阳性率分别为55.3%(21/38),18.2%(4/22),二者相比,差异具有显著性,p=0.0050; CD44v6mRNA在大肠癌组中阳性率为50.0%(19/38),在健康对照组中为9%(2/22),二者相比,差异具有显著性,p=0.0014。提示粪便中COX-2, CK20, CD44v6mRNA表达与大肠癌密切相关,进一步研究发现,三者联合检测可明显提高其阳性检出率,阳性率为86.8%(33/38),联合检测的Se,Sp,PPV,NPV及Ac分别为81.6%,81.8%,88.6%,72%及81.7%。
2.粪便基因标记物COX-2, CK20, CD44v6mRNA联合检测定性诊断肠梗阻中的应用。
对27例通过X线或腹部CT证实为肠梗阻的病人,检测粪便基因标记物mRNA表达,同时手术证实20例为癌性肠梗阻,7例为非癌性肠梗阻,20例癌性肠梗阻病人中有14例伴发肝转移,对癌性肠梗阻的诊断率为90%(18/20),其Se, Sp, PPV, NPV及Ac分别为94.7%,85.7%,90%,85.7%及88.9%。
结论：根据以上结果得出如下结论：
1.粪便COX-2, CK20, CD44v6mRNA表达与大肠癌原发灶中的表达具有同源性,说明粪便中COX-2, CK20, CD44v6mRNA来源于原发灶肿瘤细胞的脱落。
2.检测粪便中COX-2, CK20, CD44v6mRNA表达可用于结直肠癌的诊断,联合检测有助于提高其阳性检出率。
3.通过联合检测粪便COX-2, CK20, CD44v6mRNA表达可作为无法行镜检的癌性肠梗阻病人的无创性诊断手段。
Colorectal cancer (CRC) is one of the most frequent cancers and the leading to death of important disease of tumors. Colorectal cancer is the second common cause of cancer death in the western world, and is the most severe fatal disease among nonsmokers. In the United States, CRC accounts for 11% of all cancers, with an estimated 130200 new cases and 48100 death in 2001, and in Japan, there are 85000 annual cancer registrations and 35600 death caused by CRC. Because most of patients can be treated successfully if metastasis does not occur, it is important to make an early diagnosis and early treatment. In order to reduce the mortality rate of CRC, the development of the screening test by which the cancer can be diagnosed at an early stage is necessary.
To date, the fecal occult blood test(FOBT) has been used widely as the screening test for CRC as a noninvasive way. However, three recent large-scale studies showed that the sensitivity of FOBT was low using total colonoscopy as a reference standard in all subjects. However, the sensitivity of FOBT is quiet low, (15%-35%), which implies that a substantial proportion of colorectal neoplasms may be missed. In addition, the peroxidase activity present in dietary components may lead to falsepositive results, which mandates the need for special dietary restrictions prior to stool collection. With a better understanding of molecular changes associated with CRC development, tumor derived DNA alternations in stool as a noninvasive molecular screening test for colorectal neoplasmas has becoming a "hot spot" of searching. Because genetic alterations are associated directly with the devolopment of neoplasia, they have clear advantages over indirect markers such as FOBT, the disadvantage of DNA-based stool assays is the lack of sensitivity caused by clonal heterogeneity in CRC.
RNA-based stool assays have been reported in several preliminary studies, one of which showed that CD44 variant expression in human feces could be detected in 68% of CRC patients by a combination of reverse-transcription polynerase chain reaction (RT-PCR) and southern hybridization. A group of Japanese investigators has demonstrated the potential of detecting cyclooxygenase-2(COX-2)mRNA, which is frequently overexpressed in the fecal samples patients with colorectal cancer, however, whether CK20mRNA is also detected in the stool samples of patients with colorectal cancers remains unknown, whether COX-2mRNA, CK20mRNA, and CD44v6mRNA expression in the stools samples and primary lesions of CRC have hemogenous expression remains unknown. No investigations have been conducted regarding combined detection of genetic biomarkers RNA-based stool assays could improve positive-diagnosis rate of CRC and diagnose cancer-related intestinal obstruction.
Purpose:The objective of our study is to determine expression of genetic biomarkers RNA-based stool samples originate from the continuously exfoliate tumor cells shed from the primary lesions of colorectal cancers or neoplasmas, by determining expression of COX-2, CK20 and CD44v6mRNA of the primary lesions and fecal samples of CRC, providing suitable combined RNA-based stool assays for noninvasive screening test of colorectal neoplasmas or cancers. The determination of genetic biomarkers RNA-based stool assays and assessed its usefulness as a noninvasive diagnostic tool for determining cancer-related intestinal obstruction.
Method and result:We divide the experiment into several parts:
The part 1:Expression of COX-2, CK20 and CD44v6mRNA in primary colorectal carcinoma lesions and exfoliate tumor cells of fecal samples.
We carry out the expression of COX-2, CK20 and CD44v6mRNA by using RT-PCR in 38 cases of colorectal primary lesions and exfoliate tumor cells of fecal samples, we found that the agreed positive expression rate of COX-2mRNA was 44.7%(17/38) in two groups samples, the agreed negative expression rate of COX-2mRNA was 39.5%(15/38) in them, their homogenous expression rate was 84.2%(32/38); the agreed positive expression rate of CK20mRNA was 44.7%(17/38) in the primary colorectal lesions and stool samples, the agreed negative expression rate of CK20 mRNA was 36.8%(14/38), their homogenous expression rate was 81.5%(31/38); the agreed positive and negative expression rate of CD44v6mRNA in the primary lesions and fecal samples were 50%(19/38),44.7%(17/38), respectively, the homogenous expression were statistically significantly different from hemogenous expression, p<0.001, respectively, it indicated that the expression of genetic biomarkers RNA-based samples originate from the continously exfoliated tumor cells shed from the primary lesions of colorectal cancers.
Fecal samples from 38 patients with colorectal carcinoma before radical resection and from 22 cases of normal healthes severed as controls, we determined the expression of COX-2,CK20 and CD44v6mRNA of fecal exfoliated cells by using RT-PCR assay. We found that the positive expression rate of COX-2mRNA in the group of colorectal cancer and healthy control group were 47.4%(18/38),13.6%(3/22), respectively, the rate was statistically significantly different in two groups, p=0.0083; the postive expression rate of CK20mRNA in the colorectal carcinoma group and healthy control group were 55.3%(21/38),18.2%(4/22), respectively, the rate was statistically significantly different in them, p=0.0050; the postive expression rate of CD44v6mRNA in two groups were 50.0%(19/38),9%(2/22), respectively, the rate was statistially significantly different, p=0.0014, it indicated that the expression of CK20, COX-2 and CD44v6mRNA in fecal samples was closely related to the progresssion of colorectal cancers. The further study found that the combined detection of three genetic biomarkers in fecal samples can improve significantly the positive rate of colorectal cancer, the positive rate was 86.8%(33/38), obviously superior to the positive rate of single genetic biomarkers. p<0.001, the Se, Sp, PPV, NPV and Ac of combined detection of three biomarkers were 81.6%,81.8%,88.6%,72% and 81.7%, respectively.
The part 2:Combined detection of genetic biomarkers COX-2, CK20 and CD44v6mRNA and diagnose to cancer-related intestinal obstruction.
27 patients diagnosed by X-ray or abdominal CT, while 20 patients diagnosed as cancer-related intestinal obstruction by operation. The expression of COX-2, CK20, CD44v6mRNA of fecal samples were assessed by RT-PCR assay. We found that the diagnosis rate of cancer-related intestinal obstruction patients was 90%(18/20), the Se, Sp, PPV, NPV and Ac of combined detection of three genetic biomarkers were 94.7%,85.7%,90%,85.7%and 88.9%, respectively.
Conclusion:From the above date and discussion, we can draw the following conclusion:
1.There is a high homogenous expression rate of COX-2, CK20 and CD44v6mRNA in the primary colorectal cancer lesions and fecal samples, it indicated the expression of genetic biomarkers RNA-based samples originate from the continuously exfoliated tumor cells shed from the primary lesions of colorectal cancer.
2.Combined detection of expression of COX-2, CK20 and CD44v6 mRNA appears to be useful, noninvasive biological markers for diagnosis of colorectal cancer.
3.Combined detection of tumor-derived RNA-based stool assay can be as a noninvasive molecular diagnosis assay for cancer-related intestinal instead of endoscopy.
引文
[1]郝炳富,郝桂娟.结肠癌的诊断新进展.中国现代药物应用.2009，3：192-193.
[2]Kinzler KW, Vigelstein B. Lessons from hereditary colorectal cancer[J].Cell,1996,87(2):159-170
[3]Winawer SJ, Fletcher RH, Rex D, et al. Colorectal cancer screening and surveilance[J]. Gastroenterology,2003,124:544-560.
[4]Jorgensen OD, Kronborg O, Fenger C.A randomised study of screening for colorectal cancer using fecal occult blood testing:results after 13 years and seven biennial screening rounds[J]. Gut,2002,50(1):29-32.
[5]Mandel JS, Church TR, Ederer F, et al. Colorectal cancer morlality: effectiveness of biennial screening for fecal occult blood[J]. J Matl Cancer Inst,1999,91:434-437.
[6]Young GP, St, John JB, Winawer SJ, et al. Choice of fecal occult blood tests for colorectal cancer screening:recommendations based on performance characteristics in population studies.A WHO and OMED report[J]. Am J Gastroenteral,2002,97:2449-2507.
[7]Selby JV, Friedman GD, Quesenberry CP JR,et al. A case control study of screening sigmoidocopy and mortality from colorectal cancer[J]. N Engl J Med,1992,326:653-657.
[8]Kavangh AM, Giorannucci EL, Fuchs CS,et al. Screening endoscopy and risk of colorectal cancer in united states men[J]. Cancer Causes Control, 1998,9:455-462.
[9]Cheng JI, Wong JM, Hong CF, et al. Colorectal cancer screening in asymptomatic adults:comparison of colonoscopy, sigmoidoscopy and fecal ocult blood test[J]. J Formos Med Assocc,2002,101(10):685-690.
[10]Sung JJ, Chan FK, Leung WK, et al. Screening for colorectal cancer in chinese:comparison of FOBT, Flexible sigmoidoscopy, and colonoscopy[J]. Gastroenterology,2003,124(3):608-614.
[11]Citarda F, Tomasolli G, Capocaccia R,et al. Etticacy in standard clinical practice of colonoscopic polydectory in reducing colorectal cancer incidence[J]. Gut,2001,48:812-815.
[12]Tran DQ, Roqn L, Rim R,et al. Actual colonoscopy:what are the risks of perforation[J]. Am Surg,2001,67(9):845-847.
[13]Sieg A, Hachmoceller-Eisenbach U, Eisenbach T. Prospective eva lua tion of complications in outpatient GI endoscopy:a surgey among German gastroenterologists[J]. Gastrointest Endosc,2001,53(6):620-627.
[14]Rex DK, Rahmanni EY, Haseman JH,et al. Delative sensitivity of colonoscopy and bariun,enema for detection of colorectal cancer in linical prac tice[J]. Gastroenterology,1997,112:17-23.
[15]Pickhardt PJ, Choi JR, Hwang I,et al. Computed tumorgraphic colo ng rapy to screen for colorectal neoplasia in asymptomatic adults[J]. N Engl J Med, 2003,349:2191-2200.
[16]Raskin HF, Palmer WL, Kirsner JB. Erfoliative cytology in diagnosis of cancer of the colon[J]. Dis Colon Rectum,1999,2(1):46-57.
[17]Albaugh GP, Iyengar V, Lohani A,et al. Isolation of exfoliated colonic epichelial cells,a novel,non-invasive appoach to the study of cellular markers[J]. Int J Cancer,1992,52(3):347-350.
[18]Smith R, Cokkinides V, Eyre HJ. American cancer society guide line for the early detection of cancer[J]. CA Cancer Clin,2003,53(1):27-43.
[19]刘成霞.大肠肿瘤发生的主要基因学变化.滨州医学院学报.2001,24(5)：448.
[20]TeramotoH, Castellone MD, Malek RL,et al. Autocrine activation of an osteopontin-CD44-Rac Rathway enhances invasion and transformation By H-RasV 12[J]. Oncogence,2005,24(3):489-501.
[21]Gold P, Freedman S,Demonstration of tumor-specific antigens in hu man colonic carcinoma by immunological telerance and absorption techni qu es [J].J EXP Med,1965,121:439-462.
[22]Lenis J, Prevention and treatment of colorectal cancer:pay now or pay later [J]. Ann Int Med,2000,133:647-649.
[23]Anonynous. Carcinoembryonic antigen:its vole as a marker in the management of cancer.Summary of an NIH consensus statement[J]. Lancet, 1981,282:373-375.
[24]Grem J. The prognostic importance of tumor markers in adeno carcin omas of the gastrointestinal tract[J]. Curr Opinion Oneol,1997,9:380-387.
[25]Anonymous. Clinical practise guidelines for the use of tumor markers in breast and colorectal cancer[J]. Jclin Oneol,1996,14:2843-2877.
[26]Filella X, Molina R, Rique Jm, et al. CEA as a prognostic factor in Colorectal cancer[J]. Anticcancer Res,1994,14:705-708.
[27]Wagner JS, Adson MA, Van Heerden JA, et al. The natural history of hepatic metastasis from colorectal cancer:a comparision with resective treat me nt[J]. Ann Surg,1984,199:502-508.
[28]Graham RA, Wang S, Catalaro PJ,et al. Post surgical surveillance of colon cancer:meliminary cost analysis of physician examination,CEA testing. chest x-ray and colonoscopy[J]. Ann Surg,1998,228:59-63.
[29]Arnoud JP, Koehl C, Adlo M. Carcinoemhryonci antigen(CEA) in diagnosis and prognosis of coloretal carcinoma[J]. Dis Colon Rectum,1980, 23:141-144.
[30]Moertel CG, Fleming TR, Macobnald J, et al. An evaluation of the car cinoembryonic antigen(CEA) test for monitoring patients with resected colon cancer[J]. JAMA,1993,270:943-947.
[31]Rietra N, Sarli L, Costi R, et al. Pole of follow-up in management of local recurrent of coloretal cancer, a prospective randonized study [J]. Dis Colon Rectum,1998,41:1127-1133.
[32]Moll I, Cqernobilsky B, Lifschitz-Mercer B,et al. Brenner tumors but not transitional cell carcinomas of the ovary show urothelial differentiation: immuohistochemical staining of urothelial markers, including cytokeratins and wroplakins[J]. Virchows Arch,2001,438:181-191.
[33]Takano KI, Kojima T, Ogasawara N, et al. Expression of tight junction proteins in epiehelium including CK20 positive M-like cells of human adenoids in vivo and in vitro[J]. J Mol Histol,2008,39:265-273.
[34]骆成玉,李进拥.大肠癌患者门静脉血癌细胞的检测及其临床意义[J].中华外科杂志.1999,37：214.
[35]Futamura M, Takagi Y, Koumura H, et al. Spread of colorectal cancer micrometastases in regional lymph hodes by reverse transcriptase-polynerase chain rections for carcinoembryonic antigen and cytokeratin 20[J]. J Surg Oncol,1998,68:34-40.
[36]Fearon ER, Cho KR, Nigro JM, et al. Identification of a chromosome 18q gene that is altered in coloreatal cancers[J]. Science JT-science (New York,N.Y.),1990,247:49-56.
[37]Vogelstein B, Feoron ER, Hamilton SR,et al. Genetic alteration during colorectal tumor derelopment[J]. Neng LJ Med,1988,319:515-532.
[38]Cho KR, Vogelstein B, Suppressor gene alternations in the colorectal adenoma-carcinoma sequence. J cell Biochem Supppl JT-Journal of cellular biochemistry[J]. Supplcrent,1992,16G:137-141.
[39]Tanaka K, Oshimura M, Kikuchi R,et al. Suppression of tumor igenicity in colon carcinoma cells by introduction of normal chromosome 50 r18[J]. nature JT, nature,1991,349:340-342.
[40]Mehlen P, Rabizadeh S, Snipas SJ, et al. the DCC gene product indu ces apoptosis by a mechanism requiring receptor protelysis[J]. nature,1998,395: 801-804.
[41]Forcet C, Yex, Granger L, et al. the dependence receptor DCC defines an alternative mechanism for caspase activation[J]. Proc Natl Acad USA JT-proceeding of the National Academy of Science of the United States of Am erica,2001,98:3416-3421.
[42]Kim TH, Lee HK, Soo IA, et al. Notrin induces down requlation of its receptor,deleted in colorectal cancer,through the obiquitin-proteasome pathway in the embryonic cortical neuron[J]. J Neurochem JT-Journal of neuro chemi stry,2005,95:1-8.
[43]Mazelin L, Bernet A, Borod-Bidaud C, et al. Netrin-1 controls colorectal tumorigenesis by regulating apoptosis[J]. Nature JT-Nature,2004, 431:80-84.
[44]Hedrick L, Cho KR, Fdaron ER, et al. The DCC gene product in cellular differetiation and colorectal tumorigenesis [J]. Genes Devel opment 1994,8:1174-1183.
[45]Jen J, Kim H, Riantadosi S, et al. Allelic loss of chromosome 18q and prognosis in colorectal cancer[J]. N Eng LJ Med,1994,331:213-221.
[46]Kato M, Ito Y, Kobayashi S, et al. Detection of DCC and Ki-ras gene alternations in colorectal carcinoma tissue as a prognostic markers for liver metastatic recurrence[J]. Cancer JT-Cancer,1996,77:1729-1735.
[47]Martin M, Simon Assmann P, Kedinger M, et al. DCC regulates cell adhesion in human colon cancer derived HT-29 cell and associates with ezrin[J]. Eur J Cell Biol,2006,85:769-783.
[48]Shibata D, Reale MA, Lavin P, et al. The DCC protein and prognosis in coloretal cancer[J]. N Engl J Med JT-the New England Journal of Medicine, 1996,335:1727-1732.
[49]Saito M, Yamaguchi A, Goi J, et al. Expression of DCC protein in colorectal tumors and its relationship to tumor progression and metastasis [J]. Oncology JT-Oncology,1999,56:134-141.
[50]Stamenkovic I, Amiot M, Pesando JM, et al. A lymohocyte molecule implicated in lymph node homing is a member of the cartilage link protein family[J]. Cell,1989,56(3):105-107.
[51]Kim H, Yang XY, Rosada C, et al. CD44 expression in colorectal aden omas is an early event occuring prior to K-ras and P53 gene mutation[J]. Arch Biophys,1994,310(2):504-507.
[52]Wong LS, Cantrill JE,Morris AG, et al. Expression of CD44 sp lice variants in colorectal cancer[J]. Br J Surg,1997,84(6):363-367.
[53]Ishida T. Immunohistochemical expression of the CD44 variant 6 in colorectal adeno-carcinoma[J]. Surg Today,2000,30(1):28-32.
[54]Matsumura Y, Hanburg D, Smith J, et al. Non-invasive detection of m alignany by identification of usual CD44 gene activity in exfoliated cancer cells[J]. BMJ,1994,308(1):619-624.
[55]Yamao T, Matsumura Y, Shimada Y, et al. Abnormal expression of CD 44 variants in the exfoliated cells in the feces of patients with colorectal caner [J]. Gastroenterology,1998,114(6):1196-1205.
[56]Tarin D, Matsumura Y. Deranged activity of the CD44 gene and other loci as biomarkers for progression to metastatic malignancy [J]. J Cell Biochem Suppl,1993,17(1):173-185.
[57]Gunert U, Hofmann M, Dudy W, et al. A new variant of glycoprotein CD44 confers metastatic protential to vat carcinoma cells[J]. Cell,1991,65 (1):13-24.
[58]Bendardaf R, Elzagheid A, Lamlum H, et al. E-cadlerin CD44s and CD44v6 correlate with tumor differentiation in colorectal caner[J]. Oncol-Rep, 2005.13(5):831-835.
[59]Wielenga VJM, Heider KH, Otherhaus GJA, et al. Expression of CD44 variant protein in human colorectal cancer is related to tumor progres sion [J]. Caner Res,1993,53(2):4754-4756.
[60]Guo YJ, Liu GL, Wang XN, et al. Potential use of soluble CD44 in serum as indicator of tumor burden and metastasis in patients with gastric or colon cancer[J]. Cancer Res,1994,54(1):422-426.
[61]Yokoyama S, Yamane H. Predication of distant metastasis by using RT-PCR for epithelial and variant CD44mRNA in the peripheral blood of patie nts with colorectal cancer[J]. Arch Surg,2002,137(9):1069-1073.
[62]Yue SQ, Yang YL, Dou KF, et al. Expression of PCNA and CD44 m RNA in colorectal cancer with venous invasion and its relationship to liver met astasis[J]. World J Gastroenteral,2003,9(12):2863-2865.
[63]Goi T, Koneri K, Katayama K, et al. Evalvation of clinicopathological factors and the correlation between the adhesion molecule CD44 variant 9 expr ession and pulmonary metastasis from colorectal cancers[J]. Int Surg,2002, 87(2):130-136.
[64]Liu YJ, Yan PS, li J, et al. Expression and significance of CD44s、 CD44V6 and nm23mRNA in human cancer[J]. World J Gastroenteral,2005, 11(212):6601-6606.
[65]Bendardaf R, Lamlum H, Ristamaki R, et al. CD44 variant 6 expr ession predicts respense to treatment in advanced colorectal cancer[J]. Oncel Rep,2004,11(1):41-45.
[66]Vizoso FJ, Ternandez JC, Corte MD, et al. Expression and clinical significance of CD44v5 and CD44v6 in resectable colorectal cancer[J]. Cancer Res clin oncol,2004,130(11):679-686.
[67]Zavrides HN, Zizi-Sermpetzoglou A, Panousopoulos D, et al. Prog nostic evaluation of CD44 expression in correlation with bel-2 and P53 in color ectal cancer[J]. Folia Histochem Cytobiol,2005,43(1):35-36.
[68]Sano H, Kawahito Y, Wilder RL, et al. Expression of Cycloo xyge nase-1 and 2 in human colorectal cancer[J]. Cancer Res,1995,55:3785-3789.
[69]Morita I, Schindler M, Regier MK, et al. Different intracellular lo cations for prostaglandin H synthase-1 and 2[J]. J Biol Chem,1995,270(18): 10902-10908.
[70]Appleby SB, Ristimaki A, Neilson, et al. Structure of the human cyclo oxygenase-2 gene[J]. Biochem J,1994,302(pt3):723-727.
[71]Kinoshita T, Fakahashi Y, Sakashita J, et al. Growth stimulation and induction of epidemal growth factor receptor by overexpression of cycloo xy genases 1 and 2 in human colon carcinoma cells[J]. Biochem Biophysed Acta, 1999; 1438(1):120-130.
[72]Tsujii M, Kawano S, Tsuji S, et al. Cyclooxygenase regulater angio genesis induced by colon cancer cells[J]. Cell,1998; 93(5):705-716.
[73]Uefuji K, Ichikura T, et al. Cyclooxygenase-2 exression is related to prostaglandin biosynthesis and angiogenesis in human gastric [J]. Clin Cancer Res,2000,6(1):135-138.
[74]Masferrer JL, Leahy KM, et al. Antiangiogenic and antitumor Ac tiv ities of cyclooxygense-2 Inhibitors[J]. Cancer Res,2000; 60(5):1306-1311.
[75]Chen WS, Wei SJ, Liu JM, et al. Tumor invasiveness and liver meta stasis of colon cancer cells correlates with COX-2 expression and inhibited by a COX-2-selective inhibitor, etodolac[J]. Int J Cancer,2001;91(6):849-849.
[76]Zimmermann KG, Salbria M, Weber AA, et al. Cyclooxygenase-2 expression in human esophagseal carcinoma[J]. Cancer Res,1995;59:198-304.
[77]Yamauchi T, Wayanake M, Rubtoa T, et al. Cyclooxygenase-2 expre ssion as a new marker for patients with colorectal cancer[J]. Dis Colon Rectum, 2002,45:98-103.
[78]Oshima M, Dinehuk JE, Kargman SL, et al. Suppression of intestinal polyposis in delta 716 knockout mice by inhibition of colooxygenase-2 [J]. Cell, 1996;87:803-809.
[79]Masunga R, Kohno H, Dhar DK, et al. Cyclooxygenase-2 express-ion correlates with tumor neovascularzation and prognosis in human colorectal carcinoma patients[J]. Clin Cancer Res,2000,6:4064-4068.
[80]Tomozawa S, Tsuno NM, Sunarnl E, et al. Cyclooxygenase-2 over exp ression correlates with tumor recurrence especially hematogenous metas tasis of colorectal cancer[J]. Br J Cancer,2000,61:324-328.
[81]Bonenkamp JJ, Songum I, Hermans J, et al. Prognostic value of pos itive cytology findings from abdominal washing in patients with gastric canc er.Br J Surg,1996,83:612-674.
[82]Abe S, Yoshimura H, Tabata H, et al. Curative resection of gastric can cer:Limination of peritoreal lavage cytology in predicting the outcome. J Surg Oncol,1999,72:60-65.
[83]Funkai NO, Janaka J, Itami A, et al. Detection of colorectal carcinoma cells in circulating peripheral blood by RT-PCR targeting cytokeratin-20 m RNA. Life Set,1997,60:643-652.
[84]Nakanishi H, Kodera Y, Torii A, et al. Dection of carcinoembryonic antigen expressing free tumor cells on peritoreal washes from patients with gastric carcinoma by polymerase chain reaction. Ipn J Cancer Res,1997,88 (7):687-692.
[85]Nakanishi H, Kodera Y, Yamarmura Y,et al.Molecular diagnostic de tection of free cancer cells in the peritoreal cavity of patients with gas troin testinal and gynecologic maligmancies. Cancer Chemother Pharmacol,1999, 43:32-37.
[86]Nakanishi, Kodera Y, Yamarmura Y,et al. Rapid quantitative detection of free cancer cells in the peritoneal cavity of gastric cancer patients with real-time RT-PCR,and its prognostic significanse. Gan to Kagaku Ryoho,2001,28 (6):784-788.
[87]Nakanishi H, Kodera Y, Yamarmura Y,et al. Rapid quantitative dete ction of carcinoembryonci antigenexpressing free tumor cells in the peritomeal cavity of gastric cancer patients with real-time RT-PCR on the light Cycler. Int J Cancer,2000,89:411-417.
[88]于洪亮,杨茂鹏,王夫景等.大肠癌腹腔灌洗液CK20及CEAmRNA的表达意义.中国医师进修杂志,2007,30(6)：21-26.
[89]孙树印,焦华,李强.CEAmRNA,CEA蛋白及细胞学检测大肠癌腹腔液中游离癌细胞的研究.济宁医学院学报,2007,30(1)：24-26.
[90]梁俊青,李碧丽,苏秀兰.荧光定量RT-PCR技术在评估肿瘤化疗敏感性中的应用进展.内蒙古医学院学报,2010,32(3)：337-338.
[91]HUNTERK. Host genetics influence tumor metastsis[J]. Nat Rev Cancer,2006,6(2):141-146.
[92]Pinzani P, Salvadori B, Simi L, et al.Isolation by size of epithelial with breast cancer:correlation with real time reverse transcriptase-polymer-ase ch ain reaction results and feasibility of molecular analysis by laser micr odiss ection[J]. Hum Pathol,2006,37(6):711-718.
[93]Douard R, Moutereau S, Serru V, et al. Immunobead mutiplex RT-PCR detection of carcinoembroyonic genes expressing cells in the blood of colorectal cancer patients[J]. Clin Chem Lab Med,2005,43(2):127-132.
[94]Lloyel JM, Mclver CM, Stephenson SA, et al. Identification of early-stage colorectal cancer patients at risk of relapse post-resection by smmu nob ead reverse transcription-PCR analysis of peritoreal lavage fluid for malig nent cells[J]. Clin Cancer Res,2006,12(2):417-423.
[95]Ko Y, Klinz M, Totzke G, et al. Limitations of the RT-PCR for the detection of carcinoembryonic antigen-positive tumor cells in peripheral blo od [J]. Clin Caner Res,1998,4(9):2141-2146.
[96]Kopreski MS, Benko FA, Kwak LW, et al. Detection of tumor mess enger RNA in the serum of patients with malignant melanoma[J]. Clin Cancer Res,1999.5(8):1961-1965.
[97]Gradilone A, Gazzaniga P, Silvestti I, et al. Detection of CK20, CK1P and EGFRmRNA in peripheral blood of carcinoma patients correlation with clinical stsge of disease[J]. Oncol Rep,2003,10(1),217-222.
[98]Hems FA, Diepstra JH, Cornelissen IM,et al,Limitations of cyto ker atin 20 RT-PCR to detect disseminated tumor cells in blood and bone marrow of patients with colorectal cancer:expression in controls and down regulation in tumor tissue[J]. Mol Pathol,2002,55(3):156-163.
[99]Clarke LE, Leitzel K, Smith J, et al. Epidermal growth factry receptor mRNA in peripheral blood of patients with pancreatic,lung and colon carc ino mas detected by RT-PCR[J]. Int J Oncol,2003,22(2):425-430.
[100]De Luca A, Rignata S, Casamassimi A, et al. Detection of circulating tumor cells in carcinoma patients by a novel epidermal growth factor receptor reverse transcription-PCR assay[J]. Clin Cancer Res,2000,6(4):1939-1444.
[101]Leitzel K, Lieu B, Curley E, et al. Dection of cancer cells in pe ripheral blood of breast cancer patients using reverse transcription-polym-erse chain reaction for epidermal growth factor recptor[J]. Clin Cancer Res,1998, 4(12):3037-3043.
[102]Hsieh JS, Lin SR, Chang MY, et al. Apc, K-ras and P53 gene muta tion in colorectal cancer patients:correlation to chinicopathologic features and postoperative surveillance [J]. Am Surg,2007,71(4):336-343.
[103]Hibi K, Dobinson CR, Booker S, et al. Molecular detection of gen etic alterations in the serum of colerectal cancer patients[J]. Cancer Res,1998, 58(7):1405-1407.
[104]Hardingham JE, Hewett PJ, Sage RE, et al. Molecular detection of blood-bone epithelial cells in colorectal cancer patients and in patients with benign bowel disease[J]. Int J Cancer,2000,89(1):8-13.
[105]Shen CX, Hu LH, Xial,et al. Quantitative real-time RT-PCR detec tion for survivin,CK20 and CEA in peripheral blood of colorectal cancer patie nts[J]. Ipn J Clin Oncol,2008,38(11):770-771.
[106]Uen YH, Lu CY, Tsai HL, et al. Resistent presence of postoperative circulating tumor cells is a poor progrostic factor for patients with stage Ⅰ-Ⅲ colorectal cancer after surative resection[J]. Ann Surg Oncol,2008,15(8): 2120-2128.
[107]Wong CSC, Cheung MT, Ma BBY, et al. Isolated tumor cells and circulating CK20 mRNA in pNo colorectal cancer patients[J]. Int J Surg Pathol, 2008,16(2):119-126.
[108]Gervasoni A, Munoz RMM, Wengler GS, et al.Molecular signature detection of circulating tumor cells using a panel of selected genes[J]. Cancer Lett,2008,263(2):267-279.
[109]Yie SM, Lou B,Ye SR, et al. Detection of survivin-expressing circu lating cancer cells (CCCs) in periperal blood of patients with gastric and colo rectal cancer reveals high risks of relapse[J].Ann Surg Oncol,2008,15 (11):3073-3082.
[110]Garcia V, Garcia JM, Silva J, et al. Levels of VEGF-A mRNA in pla sma from patients with colorectal carcinoma as possible surrogate marker of Angiogenesis[J]. J Cancer Res Clin Oncol,2008,134(11):1165-1171.
[111]Lexther A, Gallegos N, Kocian G, et al.Detection and enumeration of circulating tumor cells in colorectal cancer[J].Br J Surg,1993,80(6):777-780.
[112]Jouve JL, Remontet L, Dancourt V, et al. Estimation of screening test(Hemoccult) sensitivity in colorectal cancer mass screening[J]. Br J Can-cer, 2001,84(11):1477.
[113]Kristinsson J, Nggaard K, Aadland E,et al.Screening of first degree relatives of patients operated for colorectal cancer evaluation of fecal calpr otectim vs. bemoccult Ⅱ [J]. Digestion,2001,64(2):104.
[114]Wong BC, Chan AO, Wong KW,et al.A pilot study of participati-on in faecal occult blood testing and screening colonoscopy after health education in Hong Kong[J]. Eur J Cancer, Prev,2005,14:181-184.
[115]Winawser S, Fletcher R, Rex D, et al. Colorechlcancer screening and surveillance clinical guidelines and rationale up date based on new evidence[J]. Gastroenterology,2003,124:544-560.
[116]Vilkin A, Pozen P, Levi Z, et al. Performance characteristics and evaluation of an automated-developed an quantitative immnochemical,fecal occult blood screening test[J]. Am J Gastroenterol,2005,100:2519-2525.
[117]Morikawa T, Kato J, Yamaji Y, et al. Comparison of the immuno-chemical fecal occult blood test and total colonoscopy in the asymptomatic pop ulation[J]. Gastroenterology,2005,129:422.
[118]Iyengar V, Albaugh GP, Lohani A, et al. Human stools as a source of viable colonic epithelial cells[J]. FASEBJ,1991,5:2856-2859.
[119]Ratto C, Flamini G, Sofo L, et al. Detection of oncogene mutation from neoplastic colonic cells exfoliated infeces[J]. Discolon Rectum,1996,39: 1238-1244.
[120]Loktionov A, Cell exfoliation in the human colon:myth,reality and implications for colorectal cancer screening[J]. Int J Cancer,2007,120:2281-2289.
[121]Akkiprik M, Ataizi-Celikel C, Dusunceli F, et al. Clinical signifi-cance of p53, k-ras and DCC gene alternations in the stage Ⅰ-Ⅱ colorectal cancers[J]. J Gastrointestis Liver Dis,2007,16:11-17.
[122]Roinar HN, Hofreiter M, Spaulding WG, et al. Molecular copro-scopy:dung and dict of the extinct ground sloth nothrotheriops shastensis[J]. Science,1998,281:402-406.
[123]Bader GM, Papanicolaou GN, The application of cytology in the diagnosis of cancer of the rectum,sigmoid,and descending colon[j]. Cancer, 1992,5:307-314.
[124]Fearon ER, Vogelstein B. A genetic model for colorected tumor-igenesis[J]. Cell,1990,61:759-767.
[125]Albaugh GP, Iyengar V, Lohani A, et al. Isolation of exfoliated colonic epielielial cells,a nomal, non-invasive approach to the study of cellular markers[J]. Int J Cancer,1992,52:347-350.
[126]Loktionov A, O'Neill IK, Silvester KR, et al. Quantitation of DNA from exfoliatled colonocyte isolated from human stool surface as a novel noninvasive screeing test for colorectal cancer[J]. Clin Cancer Res,1998,4: 337-342.
[127]Bandaletova T, Bailey N, Binghan SA, et al. Isolation of exfoliated colonocytes from human stool as a new technique for colonic cytology [J]. APMIS,2002,110:239-246.
[128]Mizuno M, Makagawa M, Uesu T,et al.Gastro-enforology,1995, 109(3):826-831.
[129]Vogeistein B, Kinzler KW, The multistep nature of cancer. Trends Genet,1993,9:138-141.
[130]Kim H. Clinical and pathological significance of the genetic analysis in colorectal carcinoma.Korean J Gastroenterol,2004,43:275-282.
[131]Fearon ER,Vogelstein B. A gentic model for colorectal tumorige-nesis. Cell,1990,61:759-767.
[132]Bord JH. Colon polyps and cancer. Endoscopy,2004,1:16-25.
[133]Isabel GG, Victor M, Matide N, et al. Standardized approach for microsatellite instability detection in colorectal carcinoma. J Matl Cancer Inst, 2000,92:544-549.
[134]Annie OC, Russell R, Broaddus PS, et al. GPG island methylation in aberrant crypt foco of the colorectum.Am J Pathol,2002,160:1823-1830.
[135]Ahlquist Dt. Stool-based DNA tests for colorectal cancer. Clinical po rential and early results.Per Gastroenterological Disord,2002,2 (suppl):s20-s26.
[136]Ratto C, Flamini G, Sofo L, et al. Detection of oncogene mutati-on from neoplastic colonic cells exfoliated in feces[J].Dis Colon Rectum,1996,39: 1238-1244.
[137]Dong SM, Traverso G, Johnson C, et al. Detecting colorectal cancer in stool with the use of multiple genetic targets[J].J Natt Cancer Inst,2001,93: 858-865.
[138]Eguchi S, Kohara N, Komuta K, et al. Mutations of the p53 gene in the stool of patients with resectable colorectal cancer[J]. Cancer,1996,77 (8 suppl):1707-1710.
[139]Davidson NO. Genetic testing in colorectal cancer:who, when, how and why[J]. Keio J Med,2007,56:14-20.
[140]Bond JH. Colon polyps andcancer[J]. Endoscopy,2005,37:208-212.
[141]Rengucci C, Maiolo P, Saragoni L, et al. Multiple detection ofgenetic alterations in tumor and stool[J]. Clin Cancer Res,2001,7:590-593
[142]Ahlquist DA, Harrington JJ, Burgart LJ, et al. Morphometric analysis of the "mucocellular layer" overlying colorectal cancer and normal muc osa: relevance to exfoliation and stool screening[J].Hum Rathol,2000,31:51-57.
[143]Huang ZH, Li LH, Yang F, et al.Detection of aberrant methylation in fecal DNA as a molecular screening tool for colorectal cancer and precan-cerous lessions[J]. World J Gastroenterol,2007,13:950-954.
[144]赵东兵,张伟等.大肠癌组织端粒酶活性的研究[J].中华肿瘤杂志.1998,20(3)：950-954.
[145]孙保存,赵秀兰等.应用端粒酶原位杂交技术检测大肠癌端粒酶活性[J].临床与实验病理学杂志.2002,18(1)：109-110.
[146]骆玉成,赵丹宁,曲军.大肠癌患者粪便标本的端粒酶活性研究[J].中华外科杂志.2001,39(8)：580-582.
[147]Ahlquist DA, Skoletsky JE, Boynton KA, et al. Colorectal cancer screening by detection of altered human DNA in stool feasibility of a multitaget assay pannel[J]. Gastroenterology,2000,119:1219-1227.
[148]Smith G, Carey FA, Beattie J, et al. Mutations in APC,K-ras and p53-alterative genetic pathways to colorectal cancer[J]. Proc Natl Acad Sci USA,2002,99:9433-9438.
[149]Wai K, Leung, Ka Fai To, Ellen P.S, et al. Detection of hypermethylated DNA or cyclooxygenase-2 messenger RNA in fecal samples of patients with colorectal cancer or polyps. Am J Gastroenteral 2007;102:1070-1076.
[150]Yamao T, Matsumura Y, Shimada Y, et al. Abnormal expression of CD44mRNA variants in the exfoliated cells in the fecel of patients with colorectal cancer[J]. Gastroenterology 1998,144(6):1196-1205.
[151]Shigeru S, Yoshida K, Miura N, et al. Potential usefulness of detec tiong cyclooxygenase 2 messenger RNA in feles for colorectal cancer scree ning[J]. Gastroenterology.2004,127(2):422-427.
[152]Stefan Wildi, Jorg KL, Haruhisa MA, et al. Charcterization of cyt okeratin 20 expression in pancreatic and xolorectal cance[J]. Cancer Res, oct, 1999,5:2840-2847.
[153]卢培琳,季峰,彭克荣等.胃癌患者CD44mRNA的表达.浙江医学.2004,26(3)：184-186.
[154]Stephen A, Bustin, Shabab S, et al. Quantification of cytokeratin 20, carcinoembryonic antigen and guanylyl cyclase C mRNA levels in lymph nodes may not predict rreatment failure in colorectal cancer patients[J] Int J Cancer,2004,108:412-417.
[2]Kinzler KW, Vigelstein B. Lessons from hereditary colorectal cancer[J].Cell,1996,87(2):159-170
[3]Winawer SJ, Fletcher RH, Rex D, et al. Colorectal cancer screening and surveilance[J]. Gastroenterology,2003,124:544-560.
[4]Jorgensen OD, Kronborg O, Fenger C.A randomised study of screening for colorectal cancer using fecal occult blood testing:results after 13 years and seven biennial screening rounds[J]. Gut,2002,50(1):29-32.
[5]Mandel JS, Church TR, Ederer F, et al. Colorectal cancer morlality: effectiveness of biennial screening for fecal occult blood[J]. J Matl Cancer Inst,1999,91:434-437.
[6]Young GP, St, John JB, Winawer SJ, et al. Choice of fecal occult blood tests for colorectal cancer screening:recommendations based on performance characteristics in population studies.A WHO and OMED report[J]. Am J Gastroenteral,2002,97:2449-2507.
[7]Selby JV, Friedman GD, Quesenberry CP JR,et al. A case control study of screening sigmoidocopy and mortality from colorectal cancer[J]. N Engl J Med,1992,326:653-657.
[8]Kavangh AM, Giorannucci EL, Fuchs CS,et al. Screening endoscopy and risk of colorectal cancer in united states men[J]. Cancer Causes Control, 1998,9:455-462.
[9]Cheng JI, Wong JM, Hong CF, et al. Colorectal cancer screening in asymptomatic adults:comparison of colonoscopy, sigmoidoscopy and fecal ocult blood test[J]. J Formos Med Assocc,2002,101(10):685-690.
[10]Sung JJ, Chan FK, Leung WK, et al. Screening for colorectal cancer in chinese:comparison of FOBT, Flexible sigmoidoscopy, and colonoscopy[J]. Gastroenterology,2003,124(3):608-614.
[11]Citarda F, Tomasolli G, Capocaccia R,et al. Etticacy in standard clinical practice of colonoscopic polydectory in reducing colorectal cancer incidence[J]. Gut,2001,48:812-815.
[12]Tran DQ, Roqn L, Rim R,et al. Actual colonoscopy:what are the risks of perforation[J]. Am Surg,2001,67(9):845-847.
[13]Sieg A, Hachmoceller-Eisenbach U, Eisenbach T. Prospective eva lua tion of complications in outpatient GI endoscopy:a surgey among German gastroenterologists[J]. Gastrointest Endosc,2001,53(6):620-627.
[14]Rex DK, Rahmanni EY, Haseman JH,et al. Delative sensitivity of colonoscopy and bariun,enema for detection of colorectal cancer in linical prac tice[J]. Gastroenterology,1997,112:17-23.
[15]Pickhardt PJ, Choi JR, Hwang I,et al. Computed tumorgraphic colo ng rapy to screen for colorectal neoplasia in asymptomatic adults[J]. N Engl J Med, 2003,349:2191-2200.
[16]Raskin HF, Palmer WL, Kirsner JB. Erfoliative cytology in diagnosis of cancer of the colon[J]. Dis Colon Rectum,1999,2(1):46-57.
[17]Albaugh GP, Iyengar V, Lohani A,et al. Isolation of exfoliated colonic epichelial cells,a novel,non-invasive appoach to the study of cellular markers[J]. Int J Cancer,1992,52(3):347-350.
[18]Smith R, Cokkinides V, Eyre HJ. American cancer society guide line for the early detection of cancer[J]. CA Cancer Clin,2003,53(1):27-43.
[19]刘成霞.大肠肿瘤发生的主要基因学变化.滨州医学院学报.2001,24(5)：448.
[20]TeramotoH, Castellone MD, Malek RL,et al. Autocrine activation of an osteopontin-CD44-Rac Rathway enhances invasion and transformation By H-RasV 12[J]. Oncogence,2005,24(3):489-501.
[21]Gold P, Freedman S,Demonstration of tumor-specific antigens in hu man colonic carcinoma by immunological telerance and absorption techni qu es [J].J EXP Med,1965,121:439-462.
[22]Lenis J, Prevention and treatment of colorectal cancer:pay now or pay later [J]. Ann Int Med,2000,133:647-649.
[23]Anonynous. Carcinoembryonic antigen:its vole as a marker in the management of cancer.Summary of an NIH consensus statement[J]. Lancet, 1981,282:373-375.
[24]Grem J. The prognostic importance of tumor markers in adeno carcin omas of the gastrointestinal tract[J]. Curr Opinion Oneol,1997,9:380-387.
[25]Anonymous. Clinical practise guidelines for the use of tumor markers in breast and colorectal cancer[J]. Jclin Oneol,1996,14:2843-2877.
[26]Filella X, Molina R, Rique Jm, et al. CEA as a prognostic factor in Colorectal cancer[J]. Anticcancer Res,1994,14:705-708.
[27]Wagner JS, Adson MA, Van Heerden JA, et al. The natural history of hepatic metastasis from colorectal cancer:a comparision with resective treat me nt[J]. Ann Surg,1984,199:502-508.
[28]Graham RA, Wang S, Catalaro PJ,et al. Post surgical surveillance of colon cancer:meliminary cost analysis of physician examination,CEA testing. chest x-ray and colonoscopy[J]. Ann Surg,1998,228:59-63.
[29]Arnoud JP, Koehl C, Adlo M. Carcinoemhryonci antigen(CEA) in diagnosis and prognosis of coloretal carcinoma[J]. Dis Colon Rectum,1980, 23:141-144.
[30]Moertel CG, Fleming TR, Macobnald J, et al. An evaluation of the car cinoembryonic antigen(CEA) test for monitoring patients with resected colon cancer[J]. JAMA,1993,270:943-947.
[31]Rietra N, Sarli L, Costi R, et al. Pole of follow-up in management of local recurrent of coloretal cancer, a prospective randonized study [J]. Dis Colon Rectum,1998,41:1127-1133.
[32]Moll I, Cqernobilsky B, Lifschitz-Mercer B,et al. Brenner tumors but not transitional cell carcinomas of the ovary show urothelial differentiation: immuohistochemical staining of urothelial markers, including cytokeratins and wroplakins[J]. Virchows Arch,2001,438:181-191.
[33]Takano KI, Kojima T, Ogasawara N, et al. Expression of tight junction proteins in epiehelium including CK20 positive M-like cells of human adenoids in vivo and in vitro[J]. J Mol Histol,2008,39:265-273.
[34]骆成玉,李进拥.大肠癌患者门静脉血癌细胞的检测及其临床意义[J].中华外科杂志.1999,37：214.
[35]Futamura M, Takagi Y, Koumura H, et al. Spread of colorectal cancer micrometastases in regional lymph hodes by reverse transcriptase-polynerase chain rections for carcinoembryonic antigen and cytokeratin 20[J]. J Surg Oncol,1998,68:34-40.
[36]Fearon ER, Cho KR, Nigro JM, et al. Identification of a chromosome 18q gene that is altered in coloreatal cancers[J]. Science JT-science (New York,N.Y.),1990,247:49-56.
[37]Vogelstein B, Feoron ER, Hamilton SR,et al. Genetic alteration during colorectal tumor derelopment[J]. Neng LJ Med,1988,319:515-532.
[38]Cho KR, Vogelstein B, Suppressor gene alternations in the colorectal adenoma-carcinoma sequence. J cell Biochem Supppl JT-Journal of cellular biochemistry[J]. Supplcrent,1992,16G:137-141.
[39]Tanaka K, Oshimura M, Kikuchi R,et al. Suppression of tumor igenicity in colon carcinoma cells by introduction of normal chromosome 50 r18[J]. nature JT, nature,1991,349:340-342.
[40]Mehlen P, Rabizadeh S, Snipas SJ, et al. the DCC gene product indu ces apoptosis by a mechanism requiring receptor protelysis[J]. nature,1998,395: 801-804.
[41]Forcet C, Yex, Granger L, et al. the dependence receptor DCC defines an alternative mechanism for caspase activation[J]. Proc Natl Acad USA JT-proceeding of the National Academy of Science of the United States of Am erica,2001,98:3416-3421.
[42]Kim TH, Lee HK, Soo IA, et al. Notrin induces down requlation of its receptor,deleted in colorectal cancer,through the obiquitin-proteasome pathway in the embryonic cortical neuron[J]. J Neurochem JT-Journal of neuro chemi stry,2005,95:1-8.
[43]Mazelin L, Bernet A, Borod-Bidaud C, et al. Netrin-1 controls colorectal tumorigenesis by regulating apoptosis[J]. Nature JT-Nature,2004, 431:80-84.
[44]Hedrick L, Cho KR, Fdaron ER, et al. The DCC gene product in cellular differetiation and colorectal tumorigenesis [J]. Genes Devel opment 1994,8:1174-1183.
[45]Jen J, Kim H, Riantadosi S, et al. Allelic loss of chromosome 18q and prognosis in colorectal cancer[J]. N Eng LJ Med,1994,331:213-221.
[46]Kato M, Ito Y, Kobayashi S, et al. Detection of DCC and Ki-ras gene alternations in colorectal carcinoma tissue as a prognostic markers for liver metastatic recurrence[J]. Cancer JT-Cancer,1996,77:1729-1735.
[47]Martin M, Simon Assmann P, Kedinger M, et al. DCC regulates cell adhesion in human colon cancer derived HT-29 cell and associates with ezrin[J]. Eur J Cell Biol,2006,85:769-783.
[48]Shibata D, Reale MA, Lavin P, et al. The DCC protein and prognosis in coloretal cancer[J]. N Engl J Med JT-the New England Journal of Medicine, 1996,335:1727-1732.
[49]Saito M, Yamaguchi A, Goi J, et al. Expression of DCC protein in colorectal tumors and its relationship to tumor progression and metastasis [J]. Oncology JT-Oncology,1999,56:134-141.
[50]Stamenkovic I, Amiot M, Pesando JM, et al. A lymohocyte molecule implicated in lymph node homing is a member of the cartilage link protein family[J]. Cell,1989,56(3):105-107.
[51]Kim H, Yang XY, Rosada C, et al. CD44 expression in colorectal aden omas is an early event occuring prior to K-ras and P53 gene mutation[J]. Arch Biophys,1994,310(2):504-507.
[52]Wong LS, Cantrill JE,Morris AG, et al. Expression of CD44 sp lice variants in colorectal cancer[J]. Br J Surg,1997,84(6):363-367.
[53]Ishida T. Immunohistochemical expression of the CD44 variant 6 in colorectal adeno-carcinoma[J]. Surg Today,2000,30(1):28-32.
[54]Matsumura Y, Hanburg D, Smith J, et al. Non-invasive detection of m alignany by identification of usual CD44 gene activity in exfoliated cancer cells[J]. BMJ,1994,308(1):619-624.
[55]Yamao T, Matsumura Y, Shimada Y, et al. Abnormal expression of CD 44 variants in the exfoliated cells in the feces of patients with colorectal caner [J]. Gastroenterology,1998,114(6):1196-1205.
[56]Tarin D, Matsumura Y. Deranged activity of the CD44 gene and other loci as biomarkers for progression to metastatic malignancy [J]. J Cell Biochem Suppl,1993,17(1):173-185.
[57]Gunert U, Hofmann M, Dudy W, et al. A new variant of glycoprotein CD44 confers metastatic protential to vat carcinoma cells[J]. Cell,1991,65 (1):13-24.
[58]Bendardaf R, Elzagheid A, Lamlum H, et al. E-cadlerin CD44s and CD44v6 correlate with tumor differentiation in colorectal caner[J]. Oncol-Rep, 2005.13(5):831-835.
[59]Wielenga VJM, Heider KH, Otherhaus GJA, et al. Expression of CD44 variant protein in human colorectal cancer is related to tumor progres sion [J]. Caner Res,1993,53(2):4754-4756.
[60]Guo YJ, Liu GL, Wang XN, et al. Potential use of soluble CD44 in serum as indicator of tumor burden and metastasis in patients with gastric or colon cancer[J]. Cancer Res,1994,54(1):422-426.
[61]Yokoyama S, Yamane H. Predication of distant metastasis by using RT-PCR for epithelial and variant CD44mRNA in the peripheral blood of patie nts with colorectal cancer[J]. Arch Surg,2002,137(9):1069-1073.
[62]Yue SQ, Yang YL, Dou KF, et al. Expression of PCNA and CD44 m RNA in colorectal cancer with venous invasion and its relationship to liver met astasis[J]. World J Gastroenteral,2003,9(12):2863-2865.
[63]Goi T, Koneri K, Katayama K, et al. Evalvation of clinicopathological factors and the correlation between the adhesion molecule CD44 variant 9 expr ession and pulmonary metastasis from colorectal cancers[J]. Int Surg,2002, 87(2):130-136.
[64]Liu YJ, Yan PS, li J, et al. Expression and significance of CD44s、 CD44V6 and nm23mRNA in human cancer[J]. World J Gastroenteral,2005, 11(212):6601-6606.
[65]Bendardaf R, Lamlum H, Ristamaki R, et al. CD44 variant 6 expr ession predicts respense to treatment in advanced colorectal cancer[J]. Oncel Rep,2004,11(1):41-45.
[66]Vizoso FJ, Ternandez JC, Corte MD, et al. Expression and clinical significance of CD44v5 and CD44v6 in resectable colorectal cancer[J]. Cancer Res clin oncol,2004,130(11):679-686.
[67]Zavrides HN, Zizi-Sermpetzoglou A, Panousopoulos D, et al. Prog nostic evaluation of CD44 expression in correlation with bel-2 and P53 in color ectal cancer[J]. Folia Histochem Cytobiol,2005,43(1):35-36.
[68]Sano H, Kawahito Y, Wilder RL, et al. Expression of Cycloo xyge nase-1 and 2 in human colorectal cancer[J]. Cancer Res,1995,55:3785-3789.
[69]Morita I, Schindler M, Regier MK, et al. Different intracellular lo cations for prostaglandin H synthase-1 and 2[J]. J Biol Chem,1995,270(18): 10902-10908.
[70]Appleby SB, Ristimaki A, Neilson, et al. Structure of the human cyclo oxygenase-2 gene[J]. Biochem J,1994,302(pt3):723-727.
[71]Kinoshita T, Fakahashi Y, Sakashita J, et al. Growth stimulation and induction of epidemal growth factor receptor by overexpression of cycloo xy genases 1 and 2 in human colon carcinoma cells[J]. Biochem Biophysed Acta, 1999; 1438(1):120-130.
[72]Tsujii M, Kawano S, Tsuji S, et al. Cyclooxygenase regulater angio genesis induced by colon cancer cells[J]. Cell,1998; 93(5):705-716.
[73]Uefuji K, Ichikura T, et al. Cyclooxygenase-2 exression is related to prostaglandin biosynthesis and angiogenesis in human gastric [J]. Clin Cancer Res,2000,6(1):135-138.
[74]Masferrer JL, Leahy KM, et al. Antiangiogenic and antitumor Ac tiv ities of cyclooxygense-2 Inhibitors[J]. Cancer Res,2000; 60(5):1306-1311.
[75]Chen WS, Wei SJ, Liu JM, et al. Tumor invasiveness and liver meta stasis of colon cancer cells correlates with COX-2 expression and inhibited by a COX-2-selective inhibitor, etodolac[J]. Int J Cancer,2001;91(6):849-849.
[76]Zimmermann KG, Salbria M, Weber AA, et al. Cyclooxygenase-2 expression in human esophagseal carcinoma[J]. Cancer Res,1995;59:198-304.
[77]Yamauchi T, Wayanake M, Rubtoa T, et al. Cyclooxygenase-2 expre ssion as a new marker for patients with colorectal cancer[J]. Dis Colon Rectum, 2002,45:98-103.
[78]Oshima M, Dinehuk JE, Kargman SL, et al. Suppression of intestinal polyposis in delta 716 knockout mice by inhibition of colooxygenase-2 [J]. Cell, 1996;87:803-809.
[79]Masunga R, Kohno H, Dhar DK, et al. Cyclooxygenase-2 express-ion correlates with tumor neovascularzation and prognosis in human colorectal carcinoma patients[J]. Clin Cancer Res,2000,6:4064-4068.
[80]Tomozawa S, Tsuno NM, Sunarnl E, et al. Cyclooxygenase-2 over exp ression correlates with tumor recurrence especially hematogenous metas tasis of colorectal cancer[J]. Br J Cancer,2000,61:324-328.
[81]Bonenkamp JJ, Songum I, Hermans J, et al. Prognostic value of pos itive cytology findings from abdominal washing in patients with gastric canc er.Br J Surg,1996,83:612-674.
[82]Abe S, Yoshimura H, Tabata H, et al. Curative resection of gastric can cer:Limination of peritoreal lavage cytology in predicting the outcome. J Surg Oncol,1999,72:60-65.
[83]Funkai NO, Janaka J, Itami A, et al. Detection of colorectal carcinoma cells in circulating peripheral blood by RT-PCR targeting cytokeratin-20 m RNA. Life Set,1997,60:643-652.
[84]Nakanishi H, Kodera Y, Torii A, et al. Dection of carcinoembryonic antigen expressing free tumor cells on peritoreal washes from patients with gastric carcinoma by polymerase chain reaction. Ipn J Cancer Res,1997,88 (7):687-692.
[85]Nakanishi H, Kodera Y, Yamarmura Y,et al.Molecular diagnostic de tection of free cancer cells in the peritoreal cavity of patients with gas troin testinal and gynecologic maligmancies. Cancer Chemother Pharmacol,1999, 43:32-37.
[86]Nakanishi, Kodera Y, Yamarmura Y,et al. Rapid quantitative detection of free cancer cells in the peritoneal cavity of gastric cancer patients with real-time RT-PCR,and its prognostic significanse. Gan to Kagaku Ryoho,2001,28 (6):784-788.
[87]Nakanishi H, Kodera Y, Yamarmura Y,et al. Rapid quantitative dete ction of carcinoembryonci antigenexpressing free tumor cells in the peritomeal cavity of gastric cancer patients with real-time RT-PCR on the light Cycler. Int J Cancer,2000,89:411-417.
[88]于洪亮,杨茂鹏,王夫景等.大肠癌腹腔灌洗液CK20及CEAmRNA的表达意义.中国医师进修杂志,2007,30(6)：21-26.
[89]孙树印,焦华,李强.CEAmRNA,CEA蛋白及细胞学检测大肠癌腹腔液中游离癌细胞的研究.济宁医学院学报,2007,30(1)：24-26.
[90]梁俊青,李碧丽,苏秀兰.荧光定量RT-PCR技术在评估肿瘤化疗敏感性中的应用进展.内蒙古医学院学报,2010,32(3)：337-338.
[91]HUNTERK. Host genetics influence tumor metastsis[J]. Nat Rev Cancer,2006,6(2):141-146.
[92]Pinzani P, Salvadori B, Simi L, et al.Isolation by size of epithelial with breast cancer:correlation with real time reverse transcriptase-polymer-ase ch ain reaction results and feasibility of molecular analysis by laser micr odiss ection[J]. Hum Pathol,2006,37(6):711-718.
[93]Douard R, Moutereau S, Serru V, et al. Immunobead mutiplex RT-PCR detection of carcinoembroyonic genes expressing cells in the blood of colorectal cancer patients[J]. Clin Chem Lab Med,2005,43(2):127-132.
[94]Lloyel JM, Mclver CM, Stephenson SA, et al. Identification of early-stage colorectal cancer patients at risk of relapse post-resection by smmu nob ead reverse transcription-PCR analysis of peritoreal lavage fluid for malig nent cells[J]. Clin Cancer Res,2006,12(2):417-423.
[95]Ko Y, Klinz M, Totzke G, et al. Limitations of the RT-PCR for the detection of carcinoembryonic antigen-positive tumor cells in peripheral blo od [J]. Clin Caner Res,1998,4(9):2141-2146.
[96]Kopreski MS, Benko FA, Kwak LW, et al. Detection of tumor mess enger RNA in the serum of patients with malignant melanoma[J]. Clin Cancer Res,1999.5(8):1961-1965.
[97]Gradilone A, Gazzaniga P, Silvestti I, et al. Detection of CK20, CK1P and EGFRmRNA in peripheral blood of carcinoma patients correlation with clinical stsge of disease[J]. Oncol Rep,2003,10(1),217-222.
[98]Hems FA, Diepstra JH, Cornelissen IM,et al,Limitations of cyto ker atin 20 RT-PCR to detect disseminated tumor cells in blood and bone marrow of patients with colorectal cancer:expression in controls and down regulation in tumor tissue[J]. Mol Pathol,2002,55(3):156-163.
[99]Clarke LE, Leitzel K, Smith J, et al. Epidermal growth factry receptor mRNA in peripheral blood of patients with pancreatic,lung and colon carc ino mas detected by RT-PCR[J]. Int J Oncol,2003,22(2):425-430.
[100]De Luca A, Rignata S, Casamassimi A, et al. Detection of circulating tumor cells in carcinoma patients by a novel epidermal growth factor receptor reverse transcription-PCR assay[J]. Clin Cancer Res,2000,6(4):1939-1444.
[101]Leitzel K, Lieu B, Curley E, et al. Dection of cancer cells in pe ripheral blood of breast cancer patients using reverse transcription-polym-erse chain reaction for epidermal growth factor recptor[J]. Clin Cancer Res,1998, 4(12):3037-3043.
[102]Hsieh JS, Lin SR, Chang MY, et al. Apc, K-ras and P53 gene muta tion in colorectal cancer patients:correlation to chinicopathologic features and postoperative surveillance [J]. Am Surg,2007,71(4):336-343.
[103]Hibi K, Dobinson CR, Booker S, et al. Molecular detection of gen etic alterations in the serum of colerectal cancer patients[J]. Cancer Res,1998, 58(7):1405-1407.
[104]Hardingham JE, Hewett PJ, Sage RE, et al. Molecular detection of blood-bone epithelial cells in colorectal cancer patients and in patients with benign bowel disease[J]. Int J Cancer,2000,89(1):8-13.
[105]Shen CX, Hu LH, Xial,et al. Quantitative real-time RT-PCR detec tion for survivin,CK20 and CEA in peripheral blood of colorectal cancer patie nts[J]. Ipn J Clin Oncol,2008,38(11):770-771.
[106]Uen YH, Lu CY, Tsai HL, et al. Resistent presence of postoperative circulating tumor cells is a poor progrostic factor for patients with stage Ⅰ-Ⅲ colorectal cancer after surative resection[J]. Ann Surg Oncol,2008,15(8): 2120-2128.
[107]Wong CSC, Cheung MT, Ma BBY, et al. Isolated tumor cells and circulating CK20 mRNA in pNo colorectal cancer patients[J]. Int J Surg Pathol, 2008,16(2):119-126.
[108]Gervasoni A, Munoz RMM, Wengler GS, et al.Molecular signature detection of circulating tumor cells using a panel of selected genes[J]. Cancer Lett,2008,263(2):267-279.
[109]Yie SM, Lou B,Ye SR, et al. Detection of survivin-expressing circu lating cancer cells (CCCs) in periperal blood of patients with gastric and colo rectal cancer reveals high risks of relapse[J].Ann Surg Oncol,2008,15 (11):3073-3082.
[110]Garcia V, Garcia JM, Silva J, et al. Levels of VEGF-A mRNA in pla sma from patients with colorectal carcinoma as possible surrogate marker of Angiogenesis[J]. J Cancer Res Clin Oncol,2008,134(11):1165-1171.
[111]Lexther A, Gallegos N, Kocian G, et al.Detection and enumeration of circulating tumor cells in colorectal cancer[J].Br J Surg,1993,80(6):777-780.
[112]Jouve JL, Remontet L, Dancourt V, et al. Estimation of screening test(Hemoccult) sensitivity in colorectal cancer mass screening[J]. Br J Can-cer, 2001,84(11):1477.
[113]Kristinsson J, Nggaard K, Aadland E,et al.Screening of first degree relatives of patients operated for colorectal cancer evaluation of fecal calpr otectim vs. bemoccult Ⅱ [J]. Digestion,2001,64(2):104.
[114]Wong BC, Chan AO, Wong KW,et al.A pilot study of participati-on in faecal occult blood testing and screening colonoscopy after health education in Hong Kong[J]. Eur J Cancer, Prev,2005,14:181-184.
[115]Winawser S, Fletcher R, Rex D, et al. Colorechlcancer screening and surveillance clinical guidelines and rationale up date based on new evidence[J]. Gastroenterology,2003,124:544-560.
[116]Vilkin A, Pozen P, Levi Z, et al. Performance characteristics and evaluation of an automated-developed an quantitative immnochemical,fecal occult blood screening test[J]. Am J Gastroenterol,2005,100:2519-2525.
[117]Morikawa T, Kato J, Yamaji Y, et al. Comparison of the immuno-chemical fecal occult blood test and total colonoscopy in the asymptomatic pop ulation[J]. Gastroenterology,2005,129:422.
[118]Iyengar V, Albaugh GP, Lohani A, et al. Human stools as a source of viable colonic epithelial cells[J]. FASEBJ,1991,5:2856-2859.
[119]Ratto C, Flamini G, Sofo L, et al. Detection of oncogene mutation from neoplastic colonic cells exfoliated infeces[J]. Discolon Rectum,1996,39: 1238-1244.
[120]Loktionov A, Cell exfoliation in the human colon:myth,reality and implications for colorectal cancer screening[J]. Int J Cancer,2007,120:2281-2289.
[121]Akkiprik M, Ataizi-Celikel C, Dusunceli F, et al. Clinical signifi-cance of p53, k-ras and DCC gene alternations in the stage Ⅰ-Ⅱ colorectal cancers[J]. J Gastrointestis Liver Dis,2007,16:11-17.
[122]Roinar HN, Hofreiter M, Spaulding WG, et al. Molecular copro-scopy:dung and dict of the extinct ground sloth nothrotheriops shastensis[J]. Science,1998,281:402-406.
[123]Bader GM, Papanicolaou GN, The application of cytology in the diagnosis of cancer of the rectum,sigmoid,and descending colon[j]. Cancer, 1992,5:307-314.
[124]Fearon ER, Vogelstein B. A genetic model for colorected tumor-igenesis[J]. Cell,1990,61:759-767.
[125]Albaugh GP, Iyengar V, Lohani A, et al. Isolation of exfoliated colonic epielielial cells,a nomal, non-invasive approach to the study of cellular markers[J]. Int J Cancer,1992,52:347-350.
[126]Loktionov A, O'Neill IK, Silvester KR, et al. Quantitation of DNA from exfoliatled colonocyte isolated from human stool surface as a novel noninvasive screeing test for colorectal cancer[J]. Clin Cancer Res,1998,4: 337-342.
[127]Bandaletova T, Bailey N, Binghan SA, et al. Isolation of exfoliated colonocytes from human stool as a new technique for colonic cytology [J]. APMIS,2002,110:239-246.
[128]Mizuno M, Makagawa M, Uesu T,et al.Gastro-enforology,1995, 109(3):826-831.
[129]Vogeistein B, Kinzler KW, The multistep nature of cancer. Trends Genet,1993,9:138-141.
[130]Kim H. Clinical and pathological significance of the genetic analysis in colorectal carcinoma.Korean J Gastroenterol,2004,43:275-282.
[131]Fearon ER,Vogelstein B. A gentic model for colorectal tumorige-nesis. Cell,1990,61:759-767.
[132]Bord JH. Colon polyps and cancer. Endoscopy,2004,1:16-25.
[133]Isabel GG, Victor M, Matide N, et al. Standardized approach for microsatellite instability detection in colorectal carcinoma. J Matl Cancer Inst, 2000,92:544-549.
[134]Annie OC, Russell R, Broaddus PS, et al. GPG island methylation in aberrant crypt foco of the colorectum.Am J Pathol,2002,160:1823-1830.
[135]Ahlquist Dt. Stool-based DNA tests for colorectal cancer. Clinical po rential and early results.Per Gastroenterological Disord,2002,2 (suppl):s20-s26.
[136]Ratto C, Flamini G, Sofo L, et al. Detection of oncogene mutati-on from neoplastic colonic cells exfoliated in feces[J].Dis Colon Rectum,1996,39: 1238-1244.
[137]Dong SM, Traverso G, Johnson C, et al. Detecting colorectal cancer in stool with the use of multiple genetic targets[J].J Natt Cancer Inst,2001,93: 858-865.
[138]Eguchi S, Kohara N, Komuta K, et al. Mutations of the p53 gene in the stool of patients with resectable colorectal cancer[J]. Cancer,1996,77 (8 suppl):1707-1710.
[139]Davidson NO. Genetic testing in colorectal cancer:who, when, how and why[J]. Keio J Med,2007,56:14-20.
[140]Bond JH. Colon polyps andcancer[J]. Endoscopy,2005,37:208-212.
[141]Rengucci C, Maiolo P, Saragoni L, et al. Multiple detection ofgenetic alterations in tumor and stool[J]. Clin Cancer Res,2001,7:590-593
[142]Ahlquist DA, Harrington JJ, Burgart LJ, et al. Morphometric analysis of the "mucocellular layer" overlying colorectal cancer and normal muc osa: relevance to exfoliation and stool screening[J].Hum Rathol,2000,31:51-57.
[143]Huang ZH, Li LH, Yang F, et al.Detection of aberrant methylation in fecal DNA as a molecular screening tool for colorectal cancer and precan-cerous lessions[J]. World J Gastroenterol,2007,13:950-954.
[144]赵东兵,张伟等.大肠癌组织端粒酶活性的研究[J].中华肿瘤杂志.1998,20(3)：950-954.
[145]孙保存,赵秀兰等.应用端粒酶原位杂交技术检测大肠癌端粒酶活性[J].临床与实验病理学杂志.2002,18(1)：109-110.
[146]骆玉成,赵丹宁,曲军.大肠癌患者粪便标本的端粒酶活性研究[J].中华外科杂志.2001,39(8)：580-582.
[147]Ahlquist DA, Skoletsky JE, Boynton KA, et al. Colorectal cancer screening by detection of altered human DNA in stool feasibility of a multitaget assay pannel[J]. Gastroenterology,2000,119:1219-1227.
[148]Smith G, Carey FA, Beattie J, et al. Mutations in APC,K-ras and p53-alterative genetic pathways to colorectal cancer[J]. Proc Natl Acad Sci USA,2002,99:9433-9438.
[149]Wai K, Leung, Ka Fai To, Ellen P.S, et al. Detection of hypermethylated DNA or cyclooxygenase-2 messenger RNA in fecal samples of patients with colorectal cancer or polyps. Am J Gastroenteral 2007;102:1070-1076.
[150]Yamao T, Matsumura Y, Shimada Y, et al. Abnormal expression of CD44mRNA variants in the exfoliated cells in the fecel of patients with colorectal cancer[J]. Gastroenterology 1998,144(6):1196-1205.
[151]Shigeru S, Yoshida K, Miura N, et al. Potential usefulness of detec tiong cyclooxygenase 2 messenger RNA in feles for colorectal cancer scree ning[J]. Gastroenterology.2004,127(2):422-427.
[152]Stefan Wildi, Jorg KL, Haruhisa MA, et al. Charcterization of cyt okeratin 20 expression in pancreatic and xolorectal cance[J]. Cancer Res, oct, 1999,5:2840-2847.
[153]卢培琳,季峰,彭克荣等.胃癌患者CD44mRNA的表达.浙江医学.2004,26(3)：184-186.
[154]Stephen A, Bustin, Shabab S, et al. Quantification of cytokeratin 20, carcinoembryonic antigen and guanylyl cyclase C mRNA levels in lymph nodes may not predict rreatment failure in colorectal cancer patients[J] Int J Cancer,2004,108:412-417.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |