非小细胞肺癌血行微转移以及基因多态性的相关研究
|
摘要
第一部分CD44V6和CK19基因表达与非小细胞肺癌血行微转移的相关研究
目的：检测非小细胞肺癌(non-small cell lung cancer,NSCLC)癌组织和外周血中CD44V6和CK19基因的表达,探讨CD44V6和CK19基因表达与NSCLC外周血微转移的关系及其临床意义。
方法：随机选择56例NSCLC患者,应用免疫组织化学技术检测NSCLC组织中CK19、CD44V6基因表达,同时采用逆转录聚合酶链反应(RT-PCR)技术检测NSCLC外周血中CK19mRNA、CD44V6mRNA表达情况；采集20例肺部良性病变患者正常肺组织和外周血作对照。
结果：
1.CD44V6基因在NSCLC组织中表达显著高于良性病变正常肺组织(P<0.001),CD44V6高表达与TNM分期(P=0.005)、淋巴结转移(P=0.024)有关。CK19在NSCLC组织和良性病变肺组织均为高表达,与临床病理特征未见相关性,无统计学差异。
2. NSCLC患者外周血中CD44V6mRNA及CK19mRNA的阳性表达率分别为64.29%(36/56)、57.14%(32/56),均显著高于对照组(P<0.05)。CD44V6mRNA在Ⅰ、Ⅱ、Ⅲ期NSCLC中阳性表达率分别为35.71%、66.67%、86.67%,其阳性表达率与临床分期(P=0.016)、淋巴结转移(P=0.02)有关；CK19mRNA在ⅠⅡ、Ⅲ期NSCLC中阳性表达率分别为28.57%、59.30%、80.0%,其阳性表达与临床分期有关(P=0.019)。
3.CD44V6和CK19在NSCLC组织中表达与外周血中表达均呈正相关(P<0.05), NSCLC外周血中CD44V6mRNA与CK19mRNA表达成正相关(P=0.002)；CD44V6mRNA与CK19mRNA联合检测阳性率为75.0%,优于单基因检测。
结论：CD44V6和CK19基因在NSCLC癌组织和外周血中表达较高,CD44V6高表达与NSLCL的侵袭转移特性相关,CD44V6mRNA和CK19mRNA联合检测有助于提高阳性率,可作为检测NSCLC患者外周血循环肿瘤细胞的分子标志物。
第二部分手术操作对非小细胞肺癌术中血行微转移的影响
目的：探讨NSCLC肺静脉血中循环肿瘤细胞(circulating tumor cell,CTC)定量检测方法,以及手术操作对非小细胞肿癌血行微转移的影响。
对象和方法：随机选择30例可手术NSCLC患者,随机分为先结扎肺静脉组和先结扎肺动脉组,以CD44V6和CK19基因作为检测标志物,采用荧光定量逆转录聚合酶链反应(FQ-RTPCR)技术检测手术前、后期肺静脉血中,CK19mRNA和CD44V6mRNA表达情况,同时采集15例健康志愿者外周血作对照。
结果：
1.NSCLC患者肺静脉血中,CD44V6mRNA和CK19mRNA分别为7.93±1.7010.76±2.74,显著高于对照组外周血,差异有统计学意义(P<0.05)。
2.肺动脉先结扎组CD44V6mRNA和CK19mRNA在手术前、后期分别为7.92±1.97vs5.67±2.11和11.21±3.14vs8.60±4.02,肺静脉先结扎组CD44V6mRNA和CK19mRNA在手术前、后期分别为7.95±1.91vs7.74±2.10和10.60±3.15vs10.30±2.98。肺静脉先结扎组在手术前后CD44V6和CK19,两者表达均无差异性(P>0.05),肺动脉先结扎组CD44V6mRNA、CK19mRNA表达在手术后期均显著高于手术前期(P≤0.05)
结论：非小细胞肿癌患者肺静脉血中可检测到循环肿瘤细胞,手术操作可能促进了血行微转移,先结扎肺静脉后结扎肺动脉一定程度可减少血行微转移。
第三部分非小细胞肺癌细胞毒T淋巴细胞抗原-4基因多态性以及与血行微转移的相关研究
目的：探讨NSCLC患者细胞毒T淋巴细胞抗原4(Cytotoxic T-lymphocyte antigen-4, CTLA-4)基因多态性的特点以及与血行微转移的关系。
方法：采用PCR-RFLP方法检测158例NSCLC患者(包括第二部分30例NSCLC患者)和72例健康对照者CTLA-4+49A>G基因分型。
结果：
1.CTLA-4+49A>G基因多态的分布
CTLA-4+49A>G基因多态的3种基因型在肺癌NSCLC患者中分布分别为：GG型44.30%(70/158),GA型41.77%(66/158)和AA型13.92%(22/158),对照组分布为：GG型48.6%(34/72),GA型41.7%(31/72)和AA型9.7%(7/72)。NSCLC患者中携带CTLA-4+49AA基因型者频率高于对照组(P=0.046),与GG基因型相比,携带AA型基因型者的相对危险度为1.489(95%CI,0.977-2.060)
2. CTLA-4+49A>G基因多态与NSCLC临床病理特征以及血行微转移的关系
CTLA-4+49A>G多态与NSCLC患者性别、病理类型、临床分期、肿瘤标记物(CEA、CYFRA21-1)、CK19mRNA以及CD44V6mRNA表达均无显著相关(P>0.05)。
结论：CTLA-4+49A>G基因多态与NSCLC临床病理特征及血行微转移未见明显相关,CTLA-4+49AA基因型可能是NSCLC的遗传易感基因,与NSCLC的发生有关。
Part One:The Correlation Study on Expression of CD44V6and CK19Gene to Micrometastasis in Non small Cell Lung Cancer
OBJECTIVE:To investigate the expression of CD44v6and CK.19gene in tumors and peripheral bood in patients with non-small cell lung cancer (NSCLC) and its clinical significance for micrometastasis.
METHODS:Fifty-six patients with NSCLC (20cases as control) were randomly enrolled into the study. The expression of CD44v6and CK19was investigated by immunohistochemistry in cancer tissues, and Reverse Transcriptase-Polymcrase Chain Reaction (RT-PCR) was used to detect the expression of the two genes in peripheral bood.
RESULTS:
1. The expression of CD44V6in cancer tissues was significantly higher than that in normal lung tisssucs(66.07%vs.0%, P<0.001). Furthermore, its expression was correlated with clinical stage(P=0.005) and lymph node metastasis(P=0.024) for patients with NSCLC. High expression of CK19was also observed in cancer tissues for the56patients, but no statistical significance was founded when compared with the normal lung tissues in the control group (83.92%vs.90%, P=NS). There was no no correlations were observed between the expression of CK19and the clinicopathological factors in patients with NSCLC.
2. In NSCLC group, the expression of CD44V6mRNA and CK19mRNA was detected in the peripheral blood of36(64.29%) patients, and32(57.14%) patients respectively. The expression of CD44V6mRNA and CK19mRNA in the peripheral blood of NSCLC was significantly higher as compared to the control group (P<0.05).The positive rate of CD44V6mRNA expression in peripheral blood of NSCLC patients was closely correlated with the pTNM stages and the status of lymph node metastasis (P=0.016,0.02, respectively). The positive rate of mRNA expression was35.71%,66.67%and86.7%in pTNM staging I, II and Ⅲ patients, respectively. Similarly, the expression of CK19mRNA was also significantly correlated with the pathological TNM stages (P=0.019). The positive rate of CK19mRNA expression was28.57%,59.30%and80.0%in pTNM staging Ⅰ, Ⅱ and Ⅲ patients, respectively.
3. The expression of CD44V6was closely correlated with CK19in both cancer tissues and peripheral blood of NSCLC patients (P=0.014,P=0.003, respectively). Combined detection of CD44V6and CK19mRNA in peripheral blood can lead to a higher sensitivity of75.0%in patients with NSCLC.
CONCLUSION:Detection of CD44V6and CK19in both cancer tissues and peripheral bood might be proper molecular markers for peripheral circular cancer cells in patients with NSCLC. Combined measurement of the two genes can further improve the diagnostic sensitivity of NSCLC.
Part two:Impacts of Surgical Procedures for Pulmonary Lobectomy on Blood Micrometastasis of Non Small Cell Lung Cancer
Objectives:The aim of this study was to investigate the impacts of different sequences in ligating pulmonary vessels during pulmonary lobectomy on blood micrometastasis of NSCLC
Materials/Methods:Cytokeratin19(CK19) and adhesion molecule CD44V6mRNA was used as molecular markers, A total of30patients with NSCLC undergoing pulmonary lobectomy were randomly divided into two groups (15cased in each group) according to the ligating sequence of the pulmonary vessels during the procedure of lobectomy. In PA-first group, pulmonary artery was ligated first; as in PV-first group, pulmonary vein was ligated in the first hand. Fluorescent quantitative RT-PCR technique was used to detect the mRNA expressions of CK19and CD44V6in pulmonary venous blood during surgery at different time-points accordingly. ACt values were calculated. Meanwhile, the peripheral blood samples from15healthy volunteers were served as control.
Results: ACt values of CD44V6mRNA and CK19mRNA in NSCLC groups at early period during surgery were7.93±1.70and10.76±2.74, compared to9.47±1.59and13.10±3.04in the control group. The expression of CD44V6mRNA and CK19mRNA in venous blood of NSCLC patients were significantly higher as compared to the control (P<0.05). In addition, the ACt values of CD44V6mRNA and CK19mRNA at early period during surgery in PA-first group were7.92±1.97and11.21±3.14, versus5.67±2.11and8.60±4.02respectively at the late period. In PA-first group.the expressions of CD44V6mRNA and CK19mRNA at late period were both significantly higher than those at the early period(P<0.05),while neither the ACt value of CD44V6mRNA nor that of CKl9mRNA at early and late periods in PV-first group displayed statistically significant differences (7.95±1.91vs7.74±2.10and10.60±3.15vs10.30±2.98, P>0.05).
Conclusions: The surgical manipulation itself might stimulate the occurrence of blood micrometastasis, thus the ligation of pulmonary vein first during pulmonary surgery may help prevent blood micrometastasis.
Part Three: Research of CTLA-4+49A>G Polymorphism in Non Small Cell Lung Cancer and its Relationship with Blood Micrometastasis.
Objectives: It was reported that Cytotoxic T-lymphocyte antigen-4(CTLA-4). as a potential immunoregulatory molecule, can suppress anti-tumor response by down-regulating T-cell activation. And the+49A>G polymorphism of CTLA-4gene was associated with several autoimmune diseases. However, little is known about functional polymorphism of CTLA-4in NSCLC and the association with blood micrometastasis.The current section was aimed to investigate functional polymorphism of CTLA-4in NSLCL and the association with blood micrometastasis.
Methods: The CTLA-4+49A>G polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in158patients with NSCLC and72healthy controls.
RESULTS: The frequencies of CTLA-4+49GG, GA and AA were detected in44.30%,41.77%and13.92%patients with NSCLC, respectively. The frequency of the CTLA-4+49AA genotype was significantly higher in patients with NSCLC compared with the corresponding controls (P=0.046). The odds of carrying the CTLA-4+49AA genotype in NSCLC group were1.489(95%CI,0.977-2.060) as compared with the CTLA-4+49GG genotype. No significant association was observed between CTLA-4+49A>G polymorphism and clinicopathologic features of NSCLC patients including gender, histopathological type, clinical stage, tumor markers (CEA and CYFRA21-1), expressions of CK19mRNA and CD44V6mRNA (P>0.05).
CONCLUSION:The polymorphism of CTLA-4+49A>G may be the susceptible factors o f NSCLC, and the AA genotype might be one of the susceptibility genes to NSCLC.
目的：检测非小细胞肺癌(non-small cell lung cancer,NSCLC)癌组织和外周血中CD44V6和CK19基因的表达,探讨CD44V6和CK19基因表达与NSCLC外周血微转移的关系及其临床意义。
方法：随机选择56例NSCLC患者,应用免疫组织化学技术检测NSCLC组织中CK19、CD44V6基因表达,同时采用逆转录聚合酶链反应(RT-PCR)技术检测NSCLC外周血中CK19mRNA、CD44V6mRNA表达情况；采集20例肺部良性病变患者正常肺组织和外周血作对照。
结果：
1.CD44V6基因在NSCLC组织中表达显著高于良性病变正常肺组织(P<0.001),CD44V6高表达与TNM分期(P=0.005)、淋巴结转移(P=0.024)有关。CK19在NSCLC组织和良性病变肺组织均为高表达,与临床病理特征未见相关性,无统计学差异。
2. NSCLC患者外周血中CD44V6mRNA及CK19mRNA的阳性表达率分别为64.29%(36/56)、57.14%(32/56),均显著高于对照组(P<0.05)。CD44V6mRNA在Ⅰ、Ⅱ、Ⅲ期NSCLC中阳性表达率分别为35.71%、66.67%、86.67%,其阳性表达率与临床分期(P=0.016)、淋巴结转移(P=0.02)有关；CK19mRNA在ⅠⅡ、Ⅲ期NSCLC中阳性表达率分别为28.57%、59.30%、80.0%,其阳性表达与临床分期有关(P=0.019)。
3.CD44V6和CK19在NSCLC组织中表达与外周血中表达均呈正相关(P<0.05), NSCLC外周血中CD44V6mRNA与CK19mRNA表达成正相关(P=0.002)；CD44V6mRNA与CK19mRNA联合检测阳性率为75.0%,优于单基因检测。
结论：CD44V6和CK19基因在NSCLC癌组织和外周血中表达较高,CD44V6高表达与NSLCL的侵袭转移特性相关,CD44V6mRNA和CK19mRNA联合检测有助于提高阳性率,可作为检测NSCLC患者外周血循环肿瘤细胞的分子标志物。
第二部分手术操作对非小细胞肺癌术中血行微转移的影响
目的：探讨NSCLC肺静脉血中循环肿瘤细胞(circulating tumor cell,CTC)定量检测方法,以及手术操作对非小细胞肿癌血行微转移的影响。
对象和方法：随机选择30例可手术NSCLC患者,随机分为先结扎肺静脉组和先结扎肺动脉组,以CD44V6和CK19基因作为检测标志物,采用荧光定量逆转录聚合酶链反应(FQ-RTPCR)技术检测手术前、后期肺静脉血中,CK19mRNA和CD44V6mRNA表达情况,同时采集15例健康志愿者外周血作对照。
结果：
1.NSCLC患者肺静脉血中,CD44V6mRNA和CK19mRNA分别为7.93±1.7010.76±2.74,显著高于对照组外周血,差异有统计学意义(P<0.05)。
2.肺动脉先结扎组CD44V6mRNA和CK19mRNA在手术前、后期分别为7.92±1.97vs5.67±2.11和11.21±3.14vs8.60±4.02,肺静脉先结扎组CD44V6mRNA和CK19mRNA在手术前、后期分别为7.95±1.91vs7.74±2.10和10.60±3.15vs10.30±2.98。肺静脉先结扎组在手术前后CD44V6和CK19,两者表达均无差异性(P>0.05),肺动脉先结扎组CD44V6mRNA、CK19mRNA表达在手术后期均显著高于手术前期(P≤0.05)
结论：非小细胞肿癌患者肺静脉血中可检测到循环肿瘤细胞,手术操作可能促进了血行微转移,先结扎肺静脉后结扎肺动脉一定程度可减少血行微转移。
第三部分非小细胞肺癌细胞毒T淋巴细胞抗原-4基因多态性以及与血行微转移的相关研究
目的：探讨NSCLC患者细胞毒T淋巴细胞抗原4(Cytotoxic T-lymphocyte antigen-4, CTLA-4)基因多态性的特点以及与血行微转移的关系。
方法：采用PCR-RFLP方法检测158例NSCLC患者(包括第二部分30例NSCLC患者)和72例健康对照者CTLA-4+49A>G基因分型。
结果：
1.CTLA-4+49A>G基因多态的分布
CTLA-4+49A>G基因多态的3种基因型在肺癌NSCLC患者中分布分别为：GG型44.30%(70/158),GA型41.77%(66/158)和AA型13.92%(22/158),对照组分布为：GG型48.6%(34/72),GA型41.7%(31/72)和AA型9.7%(7/72)。NSCLC患者中携带CTLA-4+49AA基因型者频率高于对照组(P=0.046),与GG基因型相比,携带AA型基因型者的相对危险度为1.489(95%CI,0.977-2.060)
2. CTLA-4+49A>G基因多态与NSCLC临床病理特征以及血行微转移的关系
CTLA-4+49A>G多态与NSCLC患者性别、病理类型、临床分期、肿瘤标记物(CEA、CYFRA21-1)、CK19mRNA以及CD44V6mRNA表达均无显著相关(P>0.05)。
结论：CTLA-4+49A>G基因多态与NSCLC临床病理特征及血行微转移未见明显相关,CTLA-4+49AA基因型可能是NSCLC的遗传易感基因,与NSCLC的发生有关。
Part One:The Correlation Study on Expression of CD44V6and CK19Gene to Micrometastasis in Non small Cell Lung Cancer
OBJECTIVE:To investigate the expression of CD44v6and CK.19gene in tumors and peripheral bood in patients with non-small cell lung cancer (NSCLC) and its clinical significance for micrometastasis.
METHODS:Fifty-six patients with NSCLC (20cases as control) were randomly enrolled into the study. The expression of CD44v6and CK19was investigated by immunohistochemistry in cancer tissues, and Reverse Transcriptase-Polymcrase Chain Reaction (RT-PCR) was used to detect the expression of the two genes in peripheral bood.
RESULTS:
1. The expression of CD44V6in cancer tissues was significantly higher than that in normal lung tisssucs(66.07%vs.0%, P<0.001). Furthermore, its expression was correlated with clinical stage(P=0.005) and lymph node metastasis(P=0.024) for patients with NSCLC. High expression of CK19was also observed in cancer tissues for the56patients, but no statistical significance was founded when compared with the normal lung tissues in the control group (83.92%vs.90%, P=NS). There was no no correlations were observed between the expression of CK19and the clinicopathological factors in patients with NSCLC.
2. In NSCLC group, the expression of CD44V6mRNA and CK19mRNA was detected in the peripheral blood of36(64.29%) patients, and32(57.14%) patients respectively. The expression of CD44V6mRNA and CK19mRNA in the peripheral blood of NSCLC was significantly higher as compared to the control group (P<0.05).The positive rate of CD44V6mRNA expression in peripheral blood of NSCLC patients was closely correlated with the pTNM stages and the status of lymph node metastasis (P=0.016,0.02, respectively). The positive rate of mRNA expression was35.71%,66.67%and86.7%in pTNM staging I, II and Ⅲ patients, respectively. Similarly, the expression of CK19mRNA was also significantly correlated with the pathological TNM stages (P=0.019). The positive rate of CK19mRNA expression was28.57%,59.30%and80.0%in pTNM staging Ⅰ, Ⅱ and Ⅲ patients, respectively.
3. The expression of CD44V6was closely correlated with CK19in both cancer tissues and peripheral blood of NSCLC patients (P=0.014,P=0.003, respectively). Combined detection of CD44V6and CK19mRNA in peripheral blood can lead to a higher sensitivity of75.0%in patients with NSCLC.
CONCLUSION:Detection of CD44V6and CK19in both cancer tissues and peripheral bood might be proper molecular markers for peripheral circular cancer cells in patients with NSCLC. Combined measurement of the two genes can further improve the diagnostic sensitivity of NSCLC.
Part two:Impacts of Surgical Procedures for Pulmonary Lobectomy on Blood Micrometastasis of Non Small Cell Lung Cancer
Objectives:The aim of this study was to investigate the impacts of different sequences in ligating pulmonary vessels during pulmonary lobectomy on blood micrometastasis of NSCLC
Materials/Methods:Cytokeratin19(CK19) and adhesion molecule CD44V6mRNA was used as molecular markers, A total of30patients with NSCLC undergoing pulmonary lobectomy were randomly divided into two groups (15cased in each group) according to the ligating sequence of the pulmonary vessels during the procedure of lobectomy. In PA-first group, pulmonary artery was ligated first; as in PV-first group, pulmonary vein was ligated in the first hand. Fluorescent quantitative RT-PCR technique was used to detect the mRNA expressions of CK19and CD44V6in pulmonary venous blood during surgery at different time-points accordingly. ACt values were calculated. Meanwhile, the peripheral blood samples from15healthy volunteers were served as control.
Results: ACt values of CD44V6mRNA and CK19mRNA in NSCLC groups at early period during surgery were7.93±1.70and10.76±2.74, compared to9.47±1.59and13.10±3.04in the control group. The expression of CD44V6mRNA and CK19mRNA in venous blood of NSCLC patients were significantly higher as compared to the control (P<0.05). In addition, the ACt values of CD44V6mRNA and CK19mRNA at early period during surgery in PA-first group were7.92±1.97and11.21±3.14, versus5.67±2.11and8.60±4.02respectively at the late period. In PA-first group.the expressions of CD44V6mRNA and CK19mRNA at late period were both significantly higher than those at the early period(P<0.05),while neither the ACt value of CD44V6mRNA nor that of CKl9mRNA at early and late periods in PV-first group displayed statistically significant differences (7.95±1.91vs7.74±2.10and10.60±3.15vs10.30±2.98, P>0.05).
Conclusions: The surgical manipulation itself might stimulate the occurrence of blood micrometastasis, thus the ligation of pulmonary vein first during pulmonary surgery may help prevent blood micrometastasis.
Part Three: Research of CTLA-4+49A>G Polymorphism in Non Small Cell Lung Cancer and its Relationship with Blood Micrometastasis.
Objectives: It was reported that Cytotoxic T-lymphocyte antigen-4(CTLA-4). as a potential immunoregulatory molecule, can suppress anti-tumor response by down-regulating T-cell activation. And the+49A>G polymorphism of CTLA-4gene was associated with several autoimmune diseases. However, little is known about functional polymorphism of CTLA-4in NSCLC and the association with blood micrometastasis.The current section was aimed to investigate functional polymorphism of CTLA-4in NSLCL and the association with blood micrometastasis.
Methods: The CTLA-4+49A>G polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in158patients with NSCLC and72healthy controls.
RESULTS: The frequencies of CTLA-4+49GG, GA and AA were detected in44.30%,41.77%and13.92%patients with NSCLC, respectively. The frequency of the CTLA-4+49AA genotype was significantly higher in patients with NSCLC compared with the corresponding controls (P=0.046). The odds of carrying the CTLA-4+49AA genotype in NSCLC group were1.489(95%CI,0.977-2.060) as compared with the CTLA-4+49GG genotype. No significant association was observed between CTLA-4+49A>G polymorphism and clinicopathologic features of NSCLC patients including gender, histopathological type, clinical stage, tumor markers (CEA and CYFRA21-1), expressions of CK19mRNA and CD44V6mRNA (P>0.05).
CONCLUSION:The polymorphism of CTLA-4+49A>G may be the susceptible factors o f NSCLC, and the AA genotype might be one of the susceptibility genes to NSCLC.
引文
[1]陈竺.全国第三次死因回顾抽样调查报告[M].北京,中国协和医科大学出版社,2008：10-17.
[2]Dillman RO, Herndon J, Seagren SL,et al. Improved survival In stage III non-small-cell lung cancer:seven-year follow-up of Cancer and Leukemia Group B(CALGB) 8433 Trial. J Natl Cancer Inst,1996,88:1210-1215.
[3]Hirose T, Murata Y, Oki Y, et al. Relationship of circulating tumor cells to the effectiveness of cytotoxic chemotherapy in patients with metastatic non-small-cell lung cancer. Oncol Res,2012,20(2-3):131-137.
[4]Wallwiener M, Hartkopf AD, Baccelli I,et al.The prognostic impact of circulating tumor cells in subtypes of metastatic breast cancer. Breast Cancer Res Treat,2013,137(2):503-510.
[5]Khoja L, Backen A, Sloane R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker. Br J Cancer,2012,106(3):508-516.
[6]Kurusu Y, Yamashita J,Hayashi N,et al.The sequence of vessel ligation affects tumor release into the circulation. J Thorac Cardiovasc Surg,1998,116(1):107-113.
[7]Refaely Y, Sadetzki S,Chetrit A, et al. sequence of vessel interruption during lobectomy for non-small cell lung cancer:is it indeed important? J Thorac Cardiovasc Surg,2003,125(6):1313-1320.
[8]Okumura Y,Tanaka F, Yoneda K, et al. Circulating tumor cells in pulmonary venous blood of primary lung cancer patients.Ann Thorac Surg,2009,87(6):1669-1675.
[9]Phan GQ, Yang JC, Sherry RM, et al. Cancer regression and autoimmunity induced by cytotoxic T lymphocyte-associated antigen 4 blockade in patients with metastatic melanoma. Proc Natl Acad Sci USA 2003;100: 8372-8377.
[10]Ligers A,Teleshova N,Masterman T,et al.CTLA-4 gene expression is influenced by promoter and exon 1 polymorphisms. Genes Immun 2001,2:145-152.
[11l]Ghaderi A,Yeganeh F, Kalantari T, et al.Cytotoxic T lymphocyte antigen-4 gene in breast cancer. Breast Cancer Res Treat,2004,86:1-7.
[12]Daly AK, Cholerton S, Armstong M, et al. Genotyping for polymorphisms in xenobiotic metabolism as a predictor of disease susceptibility. Environ Health Perospect,1994,102 (19):56-59.
[13]Fasano M, Sabatini MT, Wieczorek R, et al.CD44 and its V6 spliced variant in lung tumors:a role in histogenesis[J]. Cancer,1997,80(1):34-411.
[14]Matusan K, Dordevic K, Mozetic V, et al. Expression of osteopontin and CD44 molecule in papillary renal cell tumors[J]. Pathol Oncol Res, 2005,11(2):108-113.
[15]Marhaba R, Zoller M. CD44 in cancer progression:adhension, migration and growth regulation[J]. Mol Histol.2004,35(3):211-231.
[16]Afify AM, Tate S, Durbin-Johnson B, et al. Expression of CD44s and CD44v6 in lung cancer and their correlation with prognostic factors[J]. Int J Biol Markers,2011,26(1):50-57.
[17]赵党儒,方军,杜光烨,等.CD44在肺癌中表达的临床研究.中华结核与呼吸杂志,1998,21：541-543.
[18]Tolg C,Hofmann M, Herrlich P.Splicing choice form ten variant exonsestablishes CD44 variability[J]. Nucleic Acids Res,1993,21(5): 1225.
[19]刘博,王莹,曲家骐,等.联合检测CD44V6和nm23H1在非小细胞肺癌中的表达及其意义[J].中国肿瘤临床,2005,32(14)：784-787.
[20]Situ D,Long H,Lin P, et al. Expression and prognostic relevance of CD44v6 in stage I non-small cell lung carcinoma[J]. J Cancer Res Clin Oncol,2010,136(8):1213-1219.
[21]Gunthert V, Hofmann M, Rudy W, et al. A new variant of glycoprotcin CD44 confers metastatic potential to rat carcinoma eel].Ccll,1991, 65(1):13-24.
[22]Tran TA, Kallakury BVS, Sheehan CE, et al.Exp ression of CD44 standard form and viaiant isforms in non-small cell lung cancinomas[J]. Human pat hol,1997,28 (7):809-814.
[23]Devriese LA, Bosma AJ, van de Heuvel MM, et al. Circulating tumor cell detection in advanced non-small cell lung cancer patients by mul ti-marker QPCR analysis. Lung Cancer,2012,75(2):242-247.
[24]黄进肃,董强刚,周允中,等.围手术期血循环中肺癌细胞与预后的关系[J].肿瘤学杂志,2002,8(1)：31-34.
[25]Osaki T, Oyama T, Gu CD, et al. Prognostic impact of micrometastatic tumor cells in the lymph nodes and bone marrow of patients with completely resected stage I non- small cell lung cancer [J]. J Clin Oncol,2002, 20(13):2930-9.
[26]Kazuro Sugi, Seiki Kobayashi, Ryuji Yagi, et al. Usefulness of sentinel lymph node biopsy for the detection of lymph node micrometastasis in early lung cancer[J]. Interactive CardioVascular and Thoracic Surgery,2008(7):913-915.
[27]Melfi FM, Lucchi M, Davini F, et al. Intraoperative sentinel lymph node mapping in stage Ⅰ non-small cell lung cancer:detection of micrometastases by polymerase chain reaction[J].Eur J Cardiothorac Surg,2008,34(1):181-186.
[28]董强刚,江晓丰,沙慧芳,等.外周血循环中肺癌细胞的流式细胞仪定量分析[J].肿瘤,2000,20：312-314.
[29]韩宝惠,廖美琳,江晓峰,等.董强刚肺癌外周血细胞角蛋白联合2F7/S5A单抗检测及其临床应用[J].中国肺癌杂志2002,5(2)：126-130.
[30]Chen TF, Jiang GL,Fu XL, et al. CK19 mRNA expression measured by reverse-transcription polymerase chain reaction (RT-PCR) in the peripheral blood of patients with non-small cell lung cancer treated by chemo-radiation:an independent prognostic factor[J]. Lung Cancer,2007,56(1):105-14.
[31]Ge M, Shi D,Wu Q, et al.Fluctuation of circulating tumor cells in patients with lung cancer by real-time fluorescent quantitative-PCR approach before and after radiotherapy[J]. J Cancer Res Ther,2005,1(4):221-226.
[32]邢光明,路晴,侯晓薇,等.乳腺癌微转移的CK19和CD44表达研究[J].中华普通外科杂志,2003,18(8)：491-493.
[33]李锋,任军,贾军,等.RT-PCR检测乳腺癌患者外周血表皮生长因子受体和细胞角质蛋白-19mRNA的表达[J].现代肿瘤医学,2005,13(2)：154-158.
[34]Wang DR,Chen GY,Liu XL, et al.CD44v6 in peripheral blood and bone marrow of patients with gastric cancer as micro-metastasis[J]. World J Gastroenterol,2006,12(1):36-42.
[35]李坚,王洪林,涂刚,等.CD44v6 mRNA在原发性肝癌患者外周血中检测的临床意义[J].肿瘤,2006,26(4)：335-338.
[36]Song J, Zhang DS, Zheng J.Expression of CD44v6 gene in normal human peripheral blood[J]. World J Gastroenterol,2005,11(23):3591-3594.
[37]季峰,郑淑蓓,卢佩琳,等.胃癌患者外周血单个核细胞和肿瘤组织表面分化抗原44变异体6检测的临床意义[J].中华检验医学杂志,2005,28(4)：403-405.
[38]杨华伟,刘剑仑,曹骥,等.乳腺癌患者外周血SBEM-mRNA和CD44V6-mRNA的检测及其临床意义[J].中国肿瘤临床,2005,32(7)：382-385.
[39]Peck K,Sher YP,Shih JY,et al.Detection and quantitation of circulating cancer cells in the peripheral blood of lung cancer patients [J]. Cancer Res,1998,58(13):2761-2765.
[40]魏玲,宋现让,王兴武,等.MDRI和GST-π在骨软组织肉瘤中的表达及其与化疗耐药的关系[J].中华肿瘤杂志,2006,28(6)：445-448.
[41].Turnbull RB, Jr, Kyle K, Watson FR, Watson FR, Spratt J. Cancer of the colon:the influence of the no-touch isolation technic on survival rate[J].Ann Surg 1967,166(3):420-427.
[42].Hayashi N, Egami H,Kai M, Kurusu Y, Takano S,Ogawa M.No-touch isolation technique reduces intraoperative shedding of tumor cells into the portal vein during resection of colorectal cancer[J]. Surgery,1999,125(4):369-374.
[43]Yamashita JI, Kurusu Y, Fujino N, et al. Detection of circulating tumor cells in patients with non-small cell lung cancer undergoing lobectomy by video-assisted thoracic surgery:a potential hazard for intraoperative hematogenous tumor cell dissemination[J]. J Thorac Cardiovasc Surg,2000,119(5):899-905.
[44]汪国文,王祖义,刘学刚,等.非小细胞肺癌术中外周血CEA mRNA的检测及意义[J].中国肿瘤临床,2008,35(9)：505-507.
[45]姚成才,姜杰,赵德平,等.肺癌术中肺血管结扎顺序对肺癌微转移影响的研究[J].中华肿瘤防治杂志,2010,17(20)：1641-1645.
[46]Yamashita J,Matsuo A, et al. Preoperative evidence of circulating tumor cells by means of reverse transcriptase-polymerase chain reaction for carcinoembryonic antigen messenger RNA is an independent predictor of survival in non-small cell lung cancer:a prospective study[J].J Thorac Cardiovasc Surg,2002,124(2):299-305.
[47]Yoon SO, Kim YT, Jung KC,et al.TTF-1 mRNA-positive circulating tumor cells in the peripheral blood predict poor prognosis in surgically resected non-small cell lung cancer patients[J].Lung Cancer,2011,71 (2):209-216.
[48]Ge MJ, Shi D, Wu QC,et al. Observation of circulating tumour cells in patients with non-small cell lung cancer by real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction in peroperative period[J].J Cancer Res Clin Oncol,2006,132(4):248-256.
[49]刘志东,许绍发,李云松,等.非小细胞肺癌循环肿瘤细胞的定量检测意义[J].中华胸心血管外科杂志,200925(3)：184-486.
[50]李捷,孙玉鹗,盛其美,等.肺癌术中血癌胚抗原mRNA的检测及意义[J].中华外科杂志,2005,43(2)：76-79.
[51]Amirghofran Z, Jalali SA,Hosseini SV, Vasei M, Sabayan B, Ghaderi A. Evaluation of CD44 and CD44v6 in colorectal carcinoma patients:soluble forms in relation to tumor tissue expression and metastasis[J].J Gastrointest Cancer,2008,39(1-4):73-78.
[52]Chen Y, Huang K, Li X, Lin X, Zhu Z, Wu Y. Generation of a stable anti-human CD44v6 scFv and analysis of its cancer-targeting ability in vitro[J]. Cancer Immunol Immunother,2010,59(6):933-942.
[53]Wolchok JD, Saenger Y:The mechanism of anti-CTLA-4 activity and the negative regulation of T-cell activation[J]. Oncologist 2008:13:2-9.
[54]Alegre ML, Fallarino F.Mechanisms of CTLA-4-Ig in tolerance induction[J].Curr Pharm Des,2006,12(2):149-160.
[55]Ueda H, Howson JM, Esposito L, et al. Undlien Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease[J]. Nature 2003; 423:506-511.
[56]Sun T, Zhou Y, Yang M, et al. Functional genetic variations in cytotoxic T-lymphocyte antigen 4 and susceptibility to multiple types of cancer [J]. Cancer Res 2008; 68:7025-7034.
[57]Qi P, Ruan CP, Wang H, et al. CTLA-4+49A 1 G polymorphism is associated with the risk but not with the progression of colorectal cancer in Chinese [J]. Int J Colorectal Dis 2010; 25:39-45.
[58]Zheng J, Yu X, Jiang L, et al. Association between the Cytotoxic T-lymphocyte antigen 4 8 49G 1 A polymorphism and cancer risk:a meta-analysis[J]. BMC Cancer,2010,10:522-530.
[59]Wynder EL, Hoffmann D. Smoking and lung cancer:scientific challenges and opportunities[J]. Cancer Res,1994,54:5284-5295.
[60]Hou L, Grillo P, Zhu ZZ,et al. COX1 and COX2 polymorphisms and gastriccancer risk in a Polish population[J]. Anticancer Res,2007,27 (10):4243-4247.
[61]Gao J, Ke Q, Ma FIX, et al. Functional polymorphisrns in the cyclooxygenase (COX-2)gene and risk of breast cancer in a Chinese populmion[J].J Toxieol Environ Health A,2007,70(11):908-915.
[62]Waldner MJ, Neurath MF. Colit is associated cancer:the role of T cells in tumor development[J]. Semin Immunopathol,2009,31(2):249-256.
[63]Cargill M, Altshuler D, Ireland J, et a]. Characterization of single nucletotide polymorphisms in coding regions of human genes[J]. Nature Genetics,1999,22:231-238.
[64]Metzler WJ, Bajorath J, Fenderson W, et al. Solution structure of human CTLA-4 and delineation of a CD80/CD86 binding site conserved in CD28[J].Nat Struct Biol,1997,4(7):527-536.
[65]Lee KM, Chuang E, Griffin M, et al. Molecular basis of T cell inactivation by CTLA-4 [J].Science,1998,282(5397):2263-2272.
[66]Munthe-Kaas MC, Carlsen KH, Helms PJ, et al. CTLA-4 polymorphisms i n al lergy and asthma and the TH1/TH2 paradigm[J]. J Allergy Clin Immunol, 2004,114(2):280-285.
[67]Wong YK, Chang KW,Cheng CY,et al. Association of CTLA-4 gene polymorphism with oral squamous cell carcinoma[J].J Oral Pathol Med. 2006:35:51-54.
[68]Takahashi M, Kimura A.HLA and CTLA4 polymorphisms may confer a synergistic risk in the susceptibility to Graves'disease[J].J Hum Genet,2010,55(5):323-326.
[69]Reiser H,Stadecker MJ. Costimulatory B7 molecules in the pathogenesis of infectious and autoimmune diseases [J]. N Engl J Med, 1996,335(18):1369-1375.
[70]Lee KM, Holdorf AD, Uustin ML, et al. T cell receptor signaling precedes immunological synapse formation[J]. Science,2002,295 (5559):1539.
[71]Colucci F, Bergman ML, Penha- Gonealves C, et al. Apoptos resistance of nonobese diabetic peripheral lymphocytes linked to the Idd5 diabetes susceptibility region [J]. Proc Natl Acad Sci USA,1997,94(16):8670
[72]Takara M, Kouki T,DeGroot LJ. CTLA-4 AT-repeat polymorphism reduces the inhibitory function of CTLA-4 in Graves' disease[J]. Thyroid.2003,13(12):1083-1089.
[73]Kwek SS,Dao V,Roy R,et al. Diversity of antigen-specific responses induced in vivo with CTLA-4 blockade in prostate cancer patients[J]. J Immunol,2012,189(7):3759-3766.
[74]Wang W,Yu D, Sarnaik AA,et al.J Transl Med. Biomarkers on melanoma patient T cells associated with ipilimumab[J] treatment,2012,12;(10):146-148.
[75]Danielli R, Ridolfi R, Chiarion-Sileni V, et al. Ipilimumab in pretreated patients with metastatic uveal melanoma:safety and clinical efficacy[J]. Cancer Immunol Immunother,2012,61(1):41-48.
[76]陈花,赵浩亮,王振兴,等.CTLA4-Ig对肝细胞移植大鼠免疫耐受的作用及其机制[J].中华实验外科杂志,2011,28(2)：245-247.
[77]Downey SG, Klapper JA, Smith FO, et al. Prognostic factors related to clinical responsein patients with metastatic melanomatreated by CTL-associated antigen-4 blockade[J]. Clin Cancer Res,2007,13: 6681-6688.
[78]Pittet MJ. Behavior of immune players in the tumor microenvironment[J]. Curr Opin Oncol,2009,21(1):53-59.
[79]Welsh MM, Applebaum KM, Spencer SK, et al. CTLA4 variants, UV-induced tolerance, and risk of non-melanoma skin cancer[J]. Cancer Res.2009:69:6158-6163
[80]Miki R, Hattori K, Taguchi Y, et al. Identification and characterization of coding single-nucleotide polymorphisms within human protocadherin-alpha and -beta gene clusters[J]. Gene,2005,349:1-14.
[81]Su TH, Chang TY, Lee YJ, et al. CTLA-4 gene and susceptibility to human papillomavirus-16-associated cervical squamouscel1 carcinoma in Taiwanese women[J]. Carcinogenesis.2007;28:1237-1240
[82]Wang SL, Huang JD, Lai MD, et al. Molecular basis of genetic variation in debrisoquine hydroxylation in Chinese subjects:polymorphism in RFL P and sequence of CYP2D6[J]. Clin Pharmacol Ther,1993,53:410-414.
[83]厚瑞萍.细胞毒性T淋巴细胞相关抗原4基因多态性与胃癌易感性的相关性研究.山东大学硕士论文,2009：37.
[84]李华,周翊峰,郭红燕,等.细胞毒性T淋巴细胞相关抗原4编码区49位点的基因多态性与宫颈癌遗传易感性的关系[J].中华肿瘤杂志,2011,33(9)：631-634.
[85]Bao Song, Tie Liu,Yanbing Liu,et al. CTLA-4 +49A>G polymorphism is associated with advanced non-small cell lung cancer prognosis[J]. Respiration,2011,82:439-444.
[86]Greene FL,Page DI, Fleming ID,et al.AJCC Cancer Staging Manual.6th Edition.New York:Sprillger,2002.
[87]O'Flaherty JD,Gray S.Richard D, et al. Circulating tumour cells their role in metastasis and their cl inical utility inlung cancer [J]. Lung Cancer,2012,76(1):19-25.
[88]Tanaka F, Yoneda K, Kondo N, et al. Circulating tumor cell as a diagnostic marker in primary lung cancer[J]. Clin Cancer Res,2009,15(22):6980-6986.
[89]Sun YF, Yang XR, Zhou J, et al. Circulating tumor cells:advances in detection methods, biological issues, and clinical relevance[J]. J Cancer Res Clin Oncol,2011 Aug;137(8):1151-1173.
[90]黄进肃,董强刚,包国良,等.血液中微转移癌细胞对非小细胞肺癌预后的影响.中华肿瘤杂志,2004,26(5)：294-296.
[91]Takao M, Takeda K. Enumeration, characterization, and collection of intact circulating tumor cells by cross contamination free flow cytometry[J]. Cytometry A,2011,79(2):107-117.
[92]佟倩,董秀玲,高亚杰,等.乳腺癌患者外周血CK19表达临床意义的分析.中华肿瘤防治杂志,2009,16(23)：1872-1874.
[93]Chen Y, Zou TN, Wu ZP, et al. Detection of cytokeratin 19, human mammaglobin, and carcinoembryonic antigen-positive circulating tumor cells by three-marker reverse transcription-PCR assay and its relation to clinical outcome in early breast cancer[J]. Int J Biol Markers,2010,25(2):59-68.
[94]Tunca B, Egeli U, Cecener G, et al. CK19, CK20, EGFR and HER2 status of circulating tumor cells in patients with breast cancer[J]. Tumori,2012,98(2):243-251.
[95]Krismann M, Todt B, Schroder J, et al.Low specificity of cytokeratin 19 reverse transcriptase-polymerase chain reaction analyses for detection of hematogenous lung cancer dissemination[J].J Clin Oncol,1995,13(11):2769-2775.
[96]陈廷锋,蒋国梁,张宜勤,等.非小细胞肺癌根治性化放疗前外周血CK19mRNA表达的初步研究[J].中华放射肿瘤学杂志,2007,16(1)：21-25.
[97]Maeda J,Inoue M, Nakabayashi K, et al.Rapid diagnosis of lymph node metastasis in lung cancer with loop-mediated isothermal amplification assay using carcinoembryonic antigen-mRNA. Lung Cancer,2009,65(3):324-327.
[98]周清华,宫友陵,覃扬,等.非小细胞肺癌患者淋巴结、外周血及骨髓微转移分子诊断在肺癌外科治疗中的作用[J].中国肺癌杂志,2001,4(16)：435-439.
[99]Hu XC, Chow Lw.Fine needle aspiration may shed breast cells into peripheral blood as determined by RT-PCR. Oneology,2000,59(3):217-222.
[100]Yasumoto K, osa kI T, Watanabe Y, et al. Prognostic value of cytokeratin-positive cells in the bone and lymph nodes of patlenta with resected non small cell lung cancer:a multjcenter prospective study[J]. Ann Thorac Surg,2003,76:194-201
[101]Nosotti M, osi D, Palleschi A, et al. Immunocytochemical detection of occult tumor cells in the bone marrow:prognostic impact on early stages of lung cancer[J].Eur Surg Res,2008;41(3):267-271.
[102]Hirota M, Shimada S, Yamamoto K, et al. Pancreatectomy using the no-touch isolation technique followed by extensive intraoperative peritoneal lavage to prevent cancer cell dissemination:a pilot study [J].JOP,2005,6(2):143-151.
[2]Dillman RO, Herndon J, Seagren SL,et al. Improved survival In stage III non-small-cell lung cancer:seven-year follow-up of Cancer and Leukemia Group B(CALGB) 8433 Trial. J Natl Cancer Inst,1996,88:1210-1215.
[3]Hirose T, Murata Y, Oki Y, et al. Relationship of circulating tumor cells to the effectiveness of cytotoxic chemotherapy in patients with metastatic non-small-cell lung cancer. Oncol Res,2012,20(2-3):131-137.
[4]Wallwiener M, Hartkopf AD, Baccelli I,et al.The prognostic impact of circulating tumor cells in subtypes of metastatic breast cancer. Breast Cancer Res Treat,2013,137(2):503-510.
[5]Khoja L, Backen A, Sloane R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker. Br J Cancer,2012,106(3):508-516.
[6]Kurusu Y, Yamashita J,Hayashi N,et al.The sequence of vessel ligation affects tumor release into the circulation. J Thorac Cardiovasc Surg,1998,116(1):107-113.
[7]Refaely Y, Sadetzki S,Chetrit A, et al. sequence of vessel interruption during lobectomy for non-small cell lung cancer:is it indeed important? J Thorac Cardiovasc Surg,2003,125(6):1313-1320.
[8]Okumura Y,Tanaka F, Yoneda K, et al. Circulating tumor cells in pulmonary venous blood of primary lung cancer patients.Ann Thorac Surg,2009,87(6):1669-1675.
[9]Phan GQ, Yang JC, Sherry RM, et al. Cancer regression and autoimmunity induced by cytotoxic T lymphocyte-associated antigen 4 blockade in patients with metastatic melanoma. Proc Natl Acad Sci USA 2003;100: 8372-8377.
[10]Ligers A,Teleshova N,Masterman T,et al.CTLA-4 gene expression is influenced by promoter and exon 1 polymorphisms. Genes Immun 2001,2:145-152.
[11l]Ghaderi A,Yeganeh F, Kalantari T, et al.Cytotoxic T lymphocyte antigen-4 gene in breast cancer. Breast Cancer Res Treat,2004,86:1-7.
[12]Daly AK, Cholerton S, Armstong M, et al. Genotyping for polymorphisms in xenobiotic metabolism as a predictor of disease susceptibility. Environ Health Perospect,1994,102 (19):56-59.
[13]Fasano M, Sabatini MT, Wieczorek R, et al.CD44 and its V6 spliced variant in lung tumors:a role in histogenesis[J]. Cancer,1997,80(1):34-411.
[14]Matusan K, Dordevic K, Mozetic V, et al. Expression of osteopontin and CD44 molecule in papillary renal cell tumors[J]. Pathol Oncol Res, 2005,11(2):108-113.
[15]Marhaba R, Zoller M. CD44 in cancer progression:adhension, migration and growth regulation[J]. Mol Histol.2004,35(3):211-231.
[16]Afify AM, Tate S, Durbin-Johnson B, et al. Expression of CD44s and CD44v6 in lung cancer and their correlation with prognostic factors[J]. Int J Biol Markers,2011,26(1):50-57.
[17]赵党儒,方军,杜光烨,等.CD44在肺癌中表达的临床研究.中华结核与呼吸杂志,1998,21：541-543.
[18]Tolg C,Hofmann M, Herrlich P.Splicing choice form ten variant exonsestablishes CD44 variability[J]. Nucleic Acids Res,1993,21(5): 1225.
[19]刘博,王莹,曲家骐,等.联合检测CD44V6和nm23H1在非小细胞肺癌中的表达及其意义[J].中国肿瘤临床,2005,32(14)：784-787.
[20]Situ D,Long H,Lin P, et al. Expression and prognostic relevance of CD44v6 in stage I non-small cell lung carcinoma[J]. J Cancer Res Clin Oncol,2010,136(8):1213-1219.
[21]Gunthert V, Hofmann M, Rudy W, et al. A new variant of glycoprotcin CD44 confers metastatic potential to rat carcinoma eel].Ccll,1991, 65(1):13-24.
[22]Tran TA, Kallakury BVS, Sheehan CE, et al.Exp ression of CD44 standard form and viaiant isforms in non-small cell lung cancinomas[J]. Human pat hol,1997,28 (7):809-814.
[23]Devriese LA, Bosma AJ, van de Heuvel MM, et al. Circulating tumor cell detection in advanced non-small cell lung cancer patients by mul ti-marker QPCR analysis. Lung Cancer,2012,75(2):242-247.
[24]黄进肃,董强刚,周允中,等.围手术期血循环中肺癌细胞与预后的关系[J].肿瘤学杂志,2002,8(1)：31-34.
[25]Osaki T, Oyama T, Gu CD, et al. Prognostic impact of micrometastatic tumor cells in the lymph nodes and bone marrow of patients with completely resected stage I non- small cell lung cancer [J]. J Clin Oncol,2002, 20(13):2930-9.
[26]Kazuro Sugi, Seiki Kobayashi, Ryuji Yagi, et al. Usefulness of sentinel lymph node biopsy for the detection of lymph node micrometastasis in early lung cancer[J]. Interactive CardioVascular and Thoracic Surgery,2008(7):913-915.
[27]Melfi FM, Lucchi M, Davini F, et al. Intraoperative sentinel lymph node mapping in stage Ⅰ non-small cell lung cancer:detection of micrometastases by polymerase chain reaction[J].Eur J Cardiothorac Surg,2008,34(1):181-186.
[28]董强刚,江晓丰,沙慧芳,等.外周血循环中肺癌细胞的流式细胞仪定量分析[J].肿瘤,2000,20：312-314.
[29]韩宝惠,廖美琳,江晓峰,等.董强刚肺癌外周血细胞角蛋白联合2F7/S5A单抗检测及其临床应用[J].中国肺癌杂志2002,5(2)：126-130.
[30]Chen TF, Jiang GL,Fu XL, et al. CK19 mRNA expression measured by reverse-transcription polymerase chain reaction (RT-PCR) in the peripheral blood of patients with non-small cell lung cancer treated by chemo-radiation:an independent prognostic factor[J]. Lung Cancer,2007,56(1):105-14.
[31]Ge M, Shi D,Wu Q, et al.Fluctuation of circulating tumor cells in patients with lung cancer by real-time fluorescent quantitative-PCR approach before and after radiotherapy[J]. J Cancer Res Ther,2005,1(4):221-226.
[32]邢光明,路晴,侯晓薇,等.乳腺癌微转移的CK19和CD44表达研究[J].中华普通外科杂志,2003,18(8)：491-493.
[33]李锋,任军,贾军,等.RT-PCR检测乳腺癌患者外周血表皮生长因子受体和细胞角质蛋白-19mRNA的表达[J].现代肿瘤医学,2005,13(2)：154-158.
[34]Wang DR,Chen GY,Liu XL, et al.CD44v6 in peripheral blood and bone marrow of patients with gastric cancer as micro-metastasis[J]. World J Gastroenterol,2006,12(1):36-42.
[35]李坚,王洪林,涂刚,等.CD44v6 mRNA在原发性肝癌患者外周血中检测的临床意义[J].肿瘤,2006,26(4)：335-338.
[36]Song J, Zhang DS, Zheng J.Expression of CD44v6 gene in normal human peripheral blood[J]. World J Gastroenterol,2005,11(23):3591-3594.
[37]季峰,郑淑蓓,卢佩琳,等.胃癌患者外周血单个核细胞和肿瘤组织表面分化抗原44变异体6检测的临床意义[J].中华检验医学杂志,2005,28(4)：403-405.
[38]杨华伟,刘剑仑,曹骥,等.乳腺癌患者外周血SBEM-mRNA和CD44V6-mRNA的检测及其临床意义[J].中国肿瘤临床,2005,32(7)：382-385.
[39]Peck K,Sher YP,Shih JY,et al.Detection and quantitation of circulating cancer cells in the peripheral blood of lung cancer patients [J]. Cancer Res,1998,58(13):2761-2765.
[40]魏玲,宋现让,王兴武,等.MDRI和GST-π在骨软组织肉瘤中的表达及其与化疗耐药的关系[J].中华肿瘤杂志,2006,28(6)：445-448.
[41].Turnbull RB, Jr, Kyle K, Watson FR, Watson FR, Spratt J. Cancer of the colon:the influence of the no-touch isolation technic on survival rate[J].Ann Surg 1967,166(3):420-427.
[42].Hayashi N, Egami H,Kai M, Kurusu Y, Takano S,Ogawa M.No-touch isolation technique reduces intraoperative shedding of tumor cells into the portal vein during resection of colorectal cancer[J]. Surgery,1999,125(4):369-374.
[43]Yamashita JI, Kurusu Y, Fujino N, et al. Detection of circulating tumor cells in patients with non-small cell lung cancer undergoing lobectomy by video-assisted thoracic surgery:a potential hazard for intraoperative hematogenous tumor cell dissemination[J]. J Thorac Cardiovasc Surg,2000,119(5):899-905.
[44]汪国文,王祖义,刘学刚,等.非小细胞肺癌术中外周血CEA mRNA的检测及意义[J].中国肿瘤临床,2008,35(9)：505-507.
[45]姚成才,姜杰,赵德平,等.肺癌术中肺血管结扎顺序对肺癌微转移影响的研究[J].中华肿瘤防治杂志,2010,17(20)：1641-1645.
[46]Yamashita J,Matsuo A, et al. Preoperative evidence of circulating tumor cells by means of reverse transcriptase-polymerase chain reaction for carcinoembryonic antigen messenger RNA is an independent predictor of survival in non-small cell lung cancer:a prospective study[J].J Thorac Cardiovasc Surg,2002,124(2):299-305.
[47]Yoon SO, Kim YT, Jung KC,et al.TTF-1 mRNA-positive circulating tumor cells in the peripheral blood predict poor prognosis in surgically resected non-small cell lung cancer patients[J].Lung Cancer,2011,71 (2):209-216.
[48]Ge MJ, Shi D, Wu QC,et al. Observation of circulating tumour cells in patients with non-small cell lung cancer by real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction in peroperative period[J].J Cancer Res Clin Oncol,2006,132(4):248-256.
[49]刘志东,许绍发,李云松,等.非小细胞肺癌循环肿瘤细胞的定量检测意义[J].中华胸心血管外科杂志,200925(3)：184-486.
[50]李捷,孙玉鹗,盛其美,等.肺癌术中血癌胚抗原mRNA的检测及意义[J].中华外科杂志,2005,43(2)：76-79.
[51]Amirghofran Z, Jalali SA,Hosseini SV, Vasei M, Sabayan B, Ghaderi A. Evaluation of CD44 and CD44v6 in colorectal carcinoma patients:soluble forms in relation to tumor tissue expression and metastasis[J].J Gastrointest Cancer,2008,39(1-4):73-78.
[52]Chen Y, Huang K, Li X, Lin X, Zhu Z, Wu Y. Generation of a stable anti-human CD44v6 scFv and analysis of its cancer-targeting ability in vitro[J]. Cancer Immunol Immunother,2010,59(6):933-942.
[53]Wolchok JD, Saenger Y:The mechanism of anti-CTLA-4 activity and the negative regulation of T-cell activation[J]. Oncologist 2008:13:2-9.
[54]Alegre ML, Fallarino F.Mechanisms of CTLA-4-Ig in tolerance induction[J].Curr Pharm Des,2006,12(2):149-160.
[55]Ueda H, Howson JM, Esposito L, et al. Undlien Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease[J]. Nature 2003; 423:506-511.
[56]Sun T, Zhou Y, Yang M, et al. Functional genetic variations in cytotoxic T-lymphocyte antigen 4 and susceptibility to multiple types of cancer [J]. Cancer Res 2008; 68:7025-7034.
[57]Qi P, Ruan CP, Wang H, et al. CTLA-4+49A 1 G polymorphism is associated with the risk but not with the progression of colorectal cancer in Chinese [J]. Int J Colorectal Dis 2010; 25:39-45.
[58]Zheng J, Yu X, Jiang L, et al. Association between the Cytotoxic T-lymphocyte antigen 4 8 49G 1 A polymorphism and cancer risk:a meta-analysis[J]. BMC Cancer,2010,10:522-530.
[59]Wynder EL, Hoffmann D. Smoking and lung cancer:scientific challenges and opportunities[J]. Cancer Res,1994,54:5284-5295.
[60]Hou L, Grillo P, Zhu ZZ,et al. COX1 and COX2 polymorphisms and gastriccancer risk in a Polish population[J]. Anticancer Res,2007,27 (10):4243-4247.
[61]Gao J, Ke Q, Ma FIX, et al. Functional polymorphisrns in the cyclooxygenase (COX-2)gene and risk of breast cancer in a Chinese populmion[J].J Toxieol Environ Health A,2007,70(11):908-915.
[62]Waldner MJ, Neurath MF. Colit is associated cancer:the role of T cells in tumor development[J]. Semin Immunopathol,2009,31(2):249-256.
[63]Cargill M, Altshuler D, Ireland J, et a]. Characterization of single nucletotide polymorphisms in coding regions of human genes[J]. Nature Genetics,1999,22:231-238.
[64]Metzler WJ, Bajorath J, Fenderson W, et al. Solution structure of human CTLA-4 and delineation of a CD80/CD86 binding site conserved in CD28[J].Nat Struct Biol,1997,4(7):527-536.
[65]Lee KM, Chuang E, Griffin M, et al. Molecular basis of T cell inactivation by CTLA-4 [J].Science,1998,282(5397):2263-2272.
[66]Munthe-Kaas MC, Carlsen KH, Helms PJ, et al. CTLA-4 polymorphisms i n al lergy and asthma and the TH1/TH2 paradigm[J]. J Allergy Clin Immunol, 2004,114(2):280-285.
[67]Wong YK, Chang KW,Cheng CY,et al. Association of CTLA-4 gene polymorphism with oral squamous cell carcinoma[J].J Oral Pathol Med. 2006:35:51-54.
[68]Takahashi M, Kimura A.HLA and CTLA4 polymorphisms may confer a synergistic risk in the susceptibility to Graves'disease[J].J Hum Genet,2010,55(5):323-326.
[69]Reiser H,Stadecker MJ. Costimulatory B7 molecules in the pathogenesis of infectious and autoimmune diseases [J]. N Engl J Med, 1996,335(18):1369-1375.
[70]Lee KM, Holdorf AD, Uustin ML, et al. T cell receptor signaling precedes immunological synapse formation[J]. Science,2002,295 (5559):1539.
[71]Colucci F, Bergman ML, Penha- Gonealves C, et al. Apoptos resistance of nonobese diabetic peripheral lymphocytes linked to the Idd5 diabetes susceptibility region [J]. Proc Natl Acad Sci USA,1997,94(16):8670
[72]Takara M, Kouki T,DeGroot LJ. CTLA-4 AT-repeat polymorphism reduces the inhibitory function of CTLA-4 in Graves' disease[J]. Thyroid.2003,13(12):1083-1089.
[73]Kwek SS,Dao V,Roy R,et al. Diversity of antigen-specific responses induced in vivo with CTLA-4 blockade in prostate cancer patients[J]. J Immunol,2012,189(7):3759-3766.
[74]Wang W,Yu D, Sarnaik AA,et al.J Transl Med. Biomarkers on melanoma patient T cells associated with ipilimumab[J] treatment,2012,12;(10):146-148.
[75]Danielli R, Ridolfi R, Chiarion-Sileni V, et al. Ipilimumab in pretreated patients with metastatic uveal melanoma:safety and clinical efficacy[J]. Cancer Immunol Immunother,2012,61(1):41-48.
[76]陈花,赵浩亮,王振兴,等.CTLA4-Ig对肝细胞移植大鼠免疫耐受的作用及其机制[J].中华实验外科杂志,2011,28(2)：245-247.
[77]Downey SG, Klapper JA, Smith FO, et al. Prognostic factors related to clinical responsein patients with metastatic melanomatreated by CTL-associated antigen-4 blockade[J]. Clin Cancer Res,2007,13: 6681-6688.
[78]Pittet MJ. Behavior of immune players in the tumor microenvironment[J]. Curr Opin Oncol,2009,21(1):53-59.
[79]Welsh MM, Applebaum KM, Spencer SK, et al. CTLA4 variants, UV-induced tolerance, and risk of non-melanoma skin cancer[J]. Cancer Res.2009:69:6158-6163
[80]Miki R, Hattori K, Taguchi Y, et al. Identification and characterization of coding single-nucleotide polymorphisms within human protocadherin-alpha and -beta gene clusters[J]. Gene,2005,349:1-14.
[81]Su TH, Chang TY, Lee YJ, et al. CTLA-4 gene and susceptibility to human papillomavirus-16-associated cervical squamouscel1 carcinoma in Taiwanese women[J]. Carcinogenesis.2007;28:1237-1240
[82]Wang SL, Huang JD, Lai MD, et al. Molecular basis of genetic variation in debrisoquine hydroxylation in Chinese subjects:polymorphism in RFL P and sequence of CYP2D6[J]. Clin Pharmacol Ther,1993,53:410-414.
[83]厚瑞萍.细胞毒性T淋巴细胞相关抗原4基因多态性与胃癌易感性的相关性研究.山东大学硕士论文,2009：37.
[84]李华,周翊峰,郭红燕,等.细胞毒性T淋巴细胞相关抗原4编码区49位点的基因多态性与宫颈癌遗传易感性的关系[J].中华肿瘤杂志,2011,33(9)：631-634.
[85]Bao Song, Tie Liu,Yanbing Liu,et al. CTLA-4 +49A>G polymorphism is associated with advanced non-small cell lung cancer prognosis[J]. Respiration,2011,82:439-444.
[86]Greene FL,Page DI, Fleming ID,et al.AJCC Cancer Staging Manual.6th Edition.New York:Sprillger,2002.
[87]O'Flaherty JD,Gray S.Richard D, et al. Circulating tumour cells their role in metastasis and their cl inical utility inlung cancer [J]. Lung Cancer,2012,76(1):19-25.
[88]Tanaka F, Yoneda K, Kondo N, et al. Circulating tumor cell as a diagnostic marker in primary lung cancer[J]. Clin Cancer Res,2009,15(22):6980-6986.
[89]Sun YF, Yang XR, Zhou J, et al. Circulating tumor cells:advances in detection methods, biological issues, and clinical relevance[J]. J Cancer Res Clin Oncol,2011 Aug;137(8):1151-1173.
[90]黄进肃,董强刚,包国良,等.血液中微转移癌细胞对非小细胞肺癌预后的影响.中华肿瘤杂志,2004,26(5)：294-296.
[91]Takao M, Takeda K. Enumeration, characterization, and collection of intact circulating tumor cells by cross contamination free flow cytometry[J]. Cytometry A,2011,79(2):107-117.
[92]佟倩,董秀玲,高亚杰,等.乳腺癌患者外周血CK19表达临床意义的分析.中华肿瘤防治杂志,2009,16(23)：1872-1874.
[93]Chen Y, Zou TN, Wu ZP, et al. Detection of cytokeratin 19, human mammaglobin, and carcinoembryonic antigen-positive circulating tumor cells by three-marker reverse transcription-PCR assay and its relation to clinical outcome in early breast cancer[J]. Int J Biol Markers,2010,25(2):59-68.
[94]Tunca B, Egeli U, Cecener G, et al. CK19, CK20, EGFR and HER2 status of circulating tumor cells in patients with breast cancer[J]. Tumori,2012,98(2):243-251.
[95]Krismann M, Todt B, Schroder J, et al.Low specificity of cytokeratin 19 reverse transcriptase-polymerase chain reaction analyses for detection of hematogenous lung cancer dissemination[J].J Clin Oncol,1995,13(11):2769-2775.
[96]陈廷锋,蒋国梁,张宜勤,等.非小细胞肺癌根治性化放疗前外周血CK19mRNA表达的初步研究[J].中华放射肿瘤学杂志,2007,16(1)：21-25.
[97]Maeda J,Inoue M, Nakabayashi K, et al.Rapid diagnosis of lymph node metastasis in lung cancer with loop-mediated isothermal amplification assay using carcinoembryonic antigen-mRNA. Lung Cancer,2009,65(3):324-327.
[98]周清华,宫友陵,覃扬,等.非小细胞肺癌患者淋巴结、外周血及骨髓微转移分子诊断在肺癌外科治疗中的作用[J].中国肺癌杂志,2001,4(16)：435-439.
[99]Hu XC, Chow Lw.Fine needle aspiration may shed breast cells into peripheral blood as determined by RT-PCR. Oneology,2000,59(3):217-222.
[100]Yasumoto K, osa kI T, Watanabe Y, et al. Prognostic value of cytokeratin-positive cells in the bone and lymph nodes of patlenta with resected non small cell lung cancer:a multjcenter prospective study[J]. Ann Thorac Surg,2003,76:194-201
[101]Nosotti M, osi D, Palleschi A, et al. Immunocytochemical detection of occult tumor cells in the bone marrow:prognostic impact on early stages of lung cancer[J].Eur Surg Res,2008;41(3):267-271.
[102]Hirota M, Shimada S, Yamamoto K, et al. Pancreatectomy using the no-touch isolation technique followed by extensive intraoperative peritoneal lavage to prevent cancer cell dissemination:a pilot study [J].JOP,2005,6(2):143-151.
© 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 |