高半胱氨酸61(Cyr61/CCN1)及相关分子在胃癌中的表达及其与预后的关系
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摘要
目的
胃癌(Gastric Cancer,GC)是消化道最常见的恶性肿瘤。目前临床上缺乏辅助早期诊断、指导治疗及监测预后的理想分子标志物。寻找有效的分子标志物成为胃癌研究的热点。北京肿瘤分子生物学实验室前期建立了胃癌、癌旁及正常胃黏膜组织的基因表达谱,得到一组正常胃黏膜与胃癌及癌旁组织的差异表达基因。其中高半胱氨酸61细胞外基质蛋白(cysteine-rich 61,Cyr61/CCN1)在胃癌及癌旁组织中均呈高表达,均高于正常胃黏膜组织,且Cyr61在肠型胃癌和弥漫型胃癌中的表达有差异。本研究应用半定量逆转录聚合酶链反应(RT-PCR)技术检测Cyr61在人胃癌细胞系、新鲜胃癌组织样本中的表达水平,初步验证了表达谱的结果。近年来,胃癌的发病率和死亡率逐渐下降,但青年人胃癌和食管胃交界腺癌(Adenocarcinoma of theoesophagogastric junction,AEG)患者却明显增多,且二者都有其独特的临床、病理特点,预后也较差。目前关于这两种特殊类型胃癌的深入研究还很少。因此我们从胃癌病例数据库中选择了这样两组特殊人群和类型进行Cyr61与胃癌关系的系列研究,期望能够明确Cyr61在青年人胃癌和食管胃交界腺癌的诊断、分型和/或预后判断中的作用。此外,本研究以4种人胃癌细胞系为研究对象,应用实时定量PCR(Real-Time PCR)技术检测成熟体microRNA(hsa-miR-142-5p)及其预测靶基因Cyr61表达的相关性,非常初步地探讨了可能调控Cyr61异常表达的机制。
材料方法
收集中国人民解放军总医院病理科1996年1月1日至2005年12月31日的306例胃癌病例,其中青年人胃癌128例,食管胃交界腺癌191例(13例为重复病例),制作成组织芯片;121例青年人胃癌和54例食管胃交界腺癌获得随访结果。收集10例新鲜胃癌组织标本,其中5例包括癌(T)和癌旁组织(P)及配对正常组织(N),另5例包括癌(T)和配对正常组织(N)。采用免疫组织化学染色(EnVsion二步法)检测可能与Cyr61相关的CD44、CDX2、β-catenin,PTEN,MMP9,HER1和HER2蛋白在两组胃癌中的表达差异;通过培养并提取6种人胃癌细胞系BGC823、MGC803、SGC7901、AGS、N87和MKN45,以及10例新鲜胃癌组织标本中的RNA,以RT-PCR技术检测Cyr61的表达水平;细胞免疫荧光检测Cyr61蛋白在人胃癌细胞系BGC823和MGC803中的表达及定位情况;以荧光原位杂交(Fluorescence insitu hybridization,FISH)技术检测HER2在青年人胃癌及食管胃交界腺癌中的扩增情况;在Cyr61异常表达的原因分析方面,以Real-Time PCR技术检测Cyr61及相关成熟体microRNA(hsa-miR-142-5p)在人胃癌细胞系BGC823、SGC7901、AGS和MKN45中的表达水平。采用SPSS 13.0统计软件,对上述数据进行Kaplan-Meier生存分析、COX风险比例模型构建、Spearman相关分析两变量之间的关系,设P＜0.05具有统计学意义。
结果
青年人胃癌病例的年龄分布为19～40岁,平均年龄为35.6岁,中位年龄为36岁;其中男性患者84例,女性患者44例,男女比例为1.91:1。食管胃交界腺癌病例的年龄分布为28～83岁,平均年龄为59.7岁,中位年龄为60岁;其中男性患者170例,女性患者21例,男女比例为8.1:1。青年人胃癌患者的生存时间为3.4～127.2个月,中位生存时间为24个月;食管胃交界腺癌患者的生存时间为3～86个月,中位生存时间为31个月。
影响青年人胃癌患者生存率的因素有性别,脉管癌栓,嗜酸性粒细胞浸润,肿瘤浸润深度,肿瘤最大径,淋巴结转移,远处转移和临床分期;影响食管胃交界腺癌患者生存率的因素有肿瘤浸润深度,淋巴结转移和临床分期。COX生存分析显示肿瘤浸润深度越深(P=0.0076)、伴有淋巴结转移(P=0.0368)和远处转移(P=0.0156)的青年人胃癌患者预后差;食管胃交界腺癌中,肿瘤浸润深度越深(P=0.0458),伴有淋巴结转移(P=0.0389)的患者预后差。
通过RT-PCT检测6种人胃癌细胞系,结果分析Cyr61存在差异表达,于BGC823中表达最高,AGS中最低。在新鲜配对胃癌组织中,Cyr61的mRNA在正常组织中呈低表达趋势,而在胃癌和癌旁组织中有不同程度的上调,与基因表达谱结果一致。Cyr61蛋白定位于胃癌细胞的胞浆,在青年人胃癌组中,Cyr61蛋白的表达水平与性别、WHO分型、Lauren分型、临床分期、肿瘤浸润深度、淋巴结转移和远处转移均有关,其阳性表达率随肿瘤临床分期增加而增加;在食管胃交界腺癌组中,Cyr61蛋白的表达水平与临床分期、肿瘤浸润深度有关,Cyr61蛋白阳性患者和阴性患者的预后差别有统计学意义(P=0.0001)。
在青年人胃癌组中,Cyr61和β-catenin、MMP9及HER2的表达具有相关性(P分别为0.0009、0.0000和0.0000);β-catenin阳性患者的生存率比β-catenin阴性患者的生存率低(P=0.0314),PTEN阳性患者的生存率比PTEN阴性患者的生存率高(P=0.0216),MMP9阳性患者的生存率比MMP9阴性患者的生存率低(P=0.0112);在食管胃交界腺癌组中,Cyr61和MMP9的表达具有相关性(P=0.0368),MMP9阳性患者的生存率比MMP9阴性患者的生存率低(P=0.0451)。COX生存分析显示:青年人胃癌组中,MMP9、CD44高表达的患者预后差(P分别为0.0037和0.0006);食管胃交界腺癌组中,MMP9、Cyr61高表达的患者预后差(P分别为0.0074和0.0004)。
hsa-miR-142-5p与Cyr61的相对表达量之间存在负相关性,相关系数为-0.9926(P=0.007)。
结论
1.Cyr61在胃癌细胞系BGC823,MGC803和SGC7901中高表达,在AGS中低表达;在新鲜胃癌和癌旁组织中的mRNA表达水平均高于正常胃黏膜组织。Cyr61定位于胃癌细胞的胞浆,在青年人胃癌和食管胃交界腺癌中均呈高表达。
2.青年人胃癌中,Cyr61在肠型胃癌比弥漫型胃癌表达高,其表达与临床分期呈正相关,临床分期越晚,Cyr61表达越高;Cyr61的表达与β-catenin、MMP9和HER2的表达具有相关性;单因素分析,β-catenin、MMP9阳性、PTEN阴性的患者预后比β-catenin、MMP9阴性、PTEN阳性的患者预后差。多因素分析,MMP9、CD44高表达的患者比低表达的患者预后差。
3.食管胃交界腺癌中,Cyr61的表达与临床分期呈正相关,临床分期越晚,Cyr61表达越高;Cyr61阳性表达的患者预后差。单因素分析,MMP9阳性的患者的生存率比MMP9阴性患者的生存率低。多因素分析,MMP9、Cyr61高表达的患者比低表达的患者预后差。
4.青年人胃癌的特点是弥漫型胃癌所占比例高,女性更易患弥漫型胃癌,且预后更差。单因素分析影响青年人胃癌预后的因素有性别,嗜酸性粒细胞浸润,脉管癌栓,肿瘤浸润深度,肿瘤最大径,淋巴结转移,远处转移和临床分期;多因素COX风险比例模型中,影响因素有肿瘤浸润深度,淋巴结转移和远处转移。
5.食管胃交界腺癌的特点是男性高发,组织学分型以管状腺癌最常见。单因素分析影响食管胃交界腺癌预后的因素有肿瘤浸润深度,淋巴结转移和临床分期;多因素COX风险比例模型中,肿瘤浸润深度和淋巴结转移成为入选因素。
Objective
Gastric Cancer(GC)is the most common malignant tumor of the digestive tract.At present,few molecular markers are found for early diagnosis,targeted therapy and prognosis predicting.Seeking for new clinical relevant molecular markers becomes the hotspot of GC research.The previous work of our laboratory on the gene expression profiling of GC tissue revealed that gene Cysteine-rich 61(Cyr61/CCN1)was remarkably over expressed in tumor tissue,pericancerous normal appearing tissue and normal gastric mucosa,which may play an important role in GC.In order to prove the results from the gene profiling data,we analyzed the transcription level of Cyr61 in gastric cancer cell lines,fresh tissue and the protein level of Cyr61 in 306 paraffin-embedded samples of GC(128 cases from the young age group;191 cases from the oesophagogastric junction adenocarcinoma,AEG).In recent years,though the morbility and mortality are decreasing,the incidence of GC at the young age and AEG are increasing.Little is known about these two special types of GC.Therefore,we detected the expression of Cyr61 and a panel of other markers which may relate to Cyr61 in these two special types of GC,expecting to verify the significance of Cyr61 in the prognosis of GC.Besides,the expression of microRNA(has-miR-142-Sp)which may regulate Cyr61 was detected in 4 gastric cancer cell lines.
Materials and Methods
306 cases of GC were collected and reviewed from the archives of the department of pathology in PLA General Hospital during the time period from 1996 to 2005.Tissue array were made.Cases with follow-up information were evaluated to identify valuable prognostic factors.
Envision procedure of immunohistochemistry was applied to detect the expression of Cyr61,CD44,CDX2,β-catenin,PTEN,MMP9,HER1,and HER2 on the paraffin-embedded tissue array sections.The expression of Cyr61 at the mRNA level in GC cell lines and fresh tissue samples was detected by using the methods of RT-PCR and Real time PCR.The location of Cyr61 protein was confirmed by immunofluorescence.Fluorescence in situ hybridization(FISH)was used to detect the amplification of HER2 in GC.The transcription level of microRNA in GC cell lines was detected by Real-Time PCR.The results were analyzed statistically with Kaplan-Meier plots and COX proportional hazard model by applying SPSS 13.0 software.The statistical significance level was set at P＜0.05.
Results
The mean age of GC of the young was 35.6 years with the range from 19 to 40 years.The male to female ratio was 1.91:1.The mean age of the AEG was 59.7 years with the range from 28 to 83 years.The male to female ratio was 8.1:1.121 cases of the young and 54 cases of AEG were followed-up.The longest survival time of the young patient who died of the tumor was 127.2 months,the shortest was 3.4 months.The longest survival time of the AEG patient who died of the tumor was 86 months,the shortest was 3 months.
Statistic analysis showed that in the young GC patients,vascular tumor emboli,female,eosinophile infiltration,depth of tumor invasion,large tumor diameter,lymph node metastases,distant metastasis,and late clinical stage were indicators of worse prognosis.In AEG,the depth of tumor invasion,lymph node metastasis,and late clinical stage correlated with poor outcome.The COX analysis revealed that depth of tumor invasion(P=0.0076),lymph node metastasis(P=0.0368),and distant metastasis(P=0.0156)were significantly correlated to the prognosis of GC of the young;depth of tumor invasion (P=0.0458),and lymph node metastasis(P=0.0389)significantly correlated to the prognosis of the AEG.
The highest expression of Cyr61 at the mRNA level in 6 GC cell lines was in BGC823,and the lowest was in AGS.The expression level of Cyr61 in GC tissue was much higher than it in the normal tissue.Cyr61 protein was located in cytoplasm,and its expression in the young GC patients was correlated to the WHO classification,Lauren classification,clinical stage,depth of tumor invasion, lymph node metastases,and distant metastases.The expression of Cyr61 in the AEG patients was correlated to the depth of tumor invasion,clinical stage,and shorter survival time(P=0.0001).
The expression of Cyr61 was correlated toβ-catenin,MMP9,and HER2 in young GC patients(P value=0.0009,0.0000 and 0.0000,respectively).The high expression ofβ-catenin(P=0.0314),MMP9(P=0.0112),and the loss of PTEN expression(P=0.0216)were associated with lower survival rate of young GC patients.The expression of Cyr61 was correlated to MMP9 in AEG patients (P=0.0368).The expression of MMP9(P=0.0451)was associated with lower survival rate in AEG patients.The COX analysis revealed that the high expression of MMP9 and CD44 in young GC patients and the high expression of MMP9 and Cyr61 in AEG patients were associated with lower survival rate.
There was a negative correlation between the expression of hsa-miR-142-5p and Cyr61(P=0.0007).
Conclusions
1.Cyr61 is highly expressed in GC cell lines BGC823,MGC303,and SGC7901, lowly expressed in AGS.Cyr61 mRNA expressed in GC tissue and pericancerous normal appearing tissue at a high level,which was consistent with the results of gene expression profiling.Cyr61 protein was located at the cytoplasm of GC tumor cell and was highly expressed both in young GC patients and AEG patients.
2.The expression of Cyr61 protein was more common in the intestinal type than in the diffuse type of young GC patients,and the expression had positive correlation with clinical stage,β-catenin,MMP9,and HER2.The expression ofβ-catenin,MMP9,and loss of PTEN was correlated to the low survival rate in young GC patient.The high expression of MMP9 and CD44 in young GC patients was associated with lower survival rate.
3.The expression of Cyr61 protein in AEG patients showed positive correlation to the clinical stage,and was associated with lower survival rate.MMP9 had low survival rate.The COX analysis revealed that the high expression of MMP9 and Cyr61 in AEG patients was associated with lower survival rate.
4.The main histological type of GC in young patients is diffuse type,with female predominance and worse prognosis.Vescular tumor emboli,female, eosinophile infiltration,deep tumor invasion,large tumor diameter,lymph node metastases,distant metastasis,and late clinical stage were indicators of worse prognosis.
5.The characteristics of AEG were male predominance,and with the main histological type of tubular adenocarcinoma.Deep tumor invasion,lymph node metastasis,and late clinical stage were bad indicators for prognosis.
胃癌(Gastric Cancer,GC)是消化道最常见的恶性肿瘤。目前临床上缺乏辅助早期诊断、指导治疗及监测预后的理想分子标志物。寻找有效的分子标志物成为胃癌研究的热点。北京肿瘤分子生物学实验室前期建立了胃癌、癌旁及正常胃黏膜组织的基因表达谱,得到一组正常胃黏膜与胃癌及癌旁组织的差异表达基因。其中高半胱氨酸61细胞外基质蛋白(cysteine-rich 61,Cyr61/CCN1)在胃癌及癌旁组织中均呈高表达,均高于正常胃黏膜组织,且Cyr61在肠型胃癌和弥漫型胃癌中的表达有差异。本研究应用半定量逆转录聚合酶链反应(RT-PCR)技术检测Cyr61在人胃癌细胞系、新鲜胃癌组织样本中的表达水平,初步验证了表达谱的结果。近年来,胃癌的发病率和死亡率逐渐下降,但青年人胃癌和食管胃交界腺癌(Adenocarcinoma of theoesophagogastric junction,AEG)患者却明显增多,且二者都有其独特的临床、病理特点,预后也较差。目前关于这两种特殊类型胃癌的深入研究还很少。因此我们从胃癌病例数据库中选择了这样两组特殊人群和类型进行Cyr61与胃癌关系的系列研究,期望能够明确Cyr61在青年人胃癌和食管胃交界腺癌的诊断、分型和/或预后判断中的作用。此外,本研究以4种人胃癌细胞系为研究对象,应用实时定量PCR(Real-Time PCR)技术检测成熟体microRNA(hsa-miR-142-5p)及其预测靶基因Cyr61表达的相关性,非常初步地探讨了可能调控Cyr61异常表达的机制。
材料方法
收集中国人民解放军总医院病理科1996年1月1日至2005年12月31日的306例胃癌病例,其中青年人胃癌128例,食管胃交界腺癌191例(13例为重复病例),制作成组织芯片;121例青年人胃癌和54例食管胃交界腺癌获得随访结果。收集10例新鲜胃癌组织标本,其中5例包括癌(T)和癌旁组织(P)及配对正常组织(N),另5例包括癌(T)和配对正常组织(N)。采用免疫组织化学染色(EnVsion二步法)检测可能与Cyr61相关的CD44、CDX2、β-catenin,PTEN,MMP9,HER1和HER2蛋白在两组胃癌中的表达差异;通过培养并提取6种人胃癌细胞系BGC823、MGC803、SGC7901、AGS、N87和MKN45,以及10例新鲜胃癌组织标本中的RNA,以RT-PCR技术检测Cyr61的表达水平;细胞免疫荧光检测Cyr61蛋白在人胃癌细胞系BGC823和MGC803中的表达及定位情况;以荧光原位杂交(Fluorescence insitu hybridization,FISH)技术检测HER2在青年人胃癌及食管胃交界腺癌中的扩增情况;在Cyr61异常表达的原因分析方面,以Real-Time PCR技术检测Cyr61及相关成熟体microRNA(hsa-miR-142-5p)在人胃癌细胞系BGC823、SGC7901、AGS和MKN45中的表达水平。采用SPSS 13.0统计软件,对上述数据进行Kaplan-Meier生存分析、COX风险比例模型构建、Spearman相关分析两变量之间的关系,设P＜0.05具有统计学意义。
结果
青年人胃癌病例的年龄分布为19～40岁,平均年龄为35.6岁,中位年龄为36岁;其中男性患者84例,女性患者44例,男女比例为1.91:1。食管胃交界腺癌病例的年龄分布为28～83岁,平均年龄为59.7岁,中位年龄为60岁;其中男性患者170例,女性患者21例,男女比例为8.1:1。青年人胃癌患者的生存时间为3.4～127.2个月,中位生存时间为24个月;食管胃交界腺癌患者的生存时间为3～86个月,中位生存时间为31个月。
影响青年人胃癌患者生存率的因素有性别,脉管癌栓,嗜酸性粒细胞浸润,肿瘤浸润深度,肿瘤最大径,淋巴结转移,远处转移和临床分期;影响食管胃交界腺癌患者生存率的因素有肿瘤浸润深度,淋巴结转移和临床分期。COX生存分析显示肿瘤浸润深度越深(P=0.0076)、伴有淋巴结转移(P=0.0368)和远处转移(P=0.0156)的青年人胃癌患者预后差;食管胃交界腺癌中,肿瘤浸润深度越深(P=0.0458),伴有淋巴结转移(P=0.0389)的患者预后差。
通过RT-PCT检测6种人胃癌细胞系,结果分析Cyr61存在差异表达,于BGC823中表达最高,AGS中最低。在新鲜配对胃癌组织中,Cyr61的mRNA在正常组织中呈低表达趋势,而在胃癌和癌旁组织中有不同程度的上调,与基因表达谱结果一致。Cyr61蛋白定位于胃癌细胞的胞浆,在青年人胃癌组中,Cyr61蛋白的表达水平与性别、WHO分型、Lauren分型、临床分期、肿瘤浸润深度、淋巴结转移和远处转移均有关,其阳性表达率随肿瘤临床分期增加而增加;在食管胃交界腺癌组中,Cyr61蛋白的表达水平与临床分期、肿瘤浸润深度有关,Cyr61蛋白阳性患者和阴性患者的预后差别有统计学意义(P=0.0001)。
在青年人胃癌组中,Cyr61和β-catenin、MMP9及HER2的表达具有相关性(P分别为0.0009、0.0000和0.0000);β-catenin阳性患者的生存率比β-catenin阴性患者的生存率低(P=0.0314),PTEN阳性患者的生存率比PTEN阴性患者的生存率高(P=0.0216),MMP9阳性患者的生存率比MMP9阴性患者的生存率低(P=0.0112);在食管胃交界腺癌组中,Cyr61和MMP9的表达具有相关性(P=0.0368),MMP9阳性患者的生存率比MMP9阴性患者的生存率低(P=0.0451)。COX生存分析显示:青年人胃癌组中,MMP9、CD44高表达的患者预后差(P分别为0.0037和0.0006);食管胃交界腺癌组中,MMP9、Cyr61高表达的患者预后差(P分别为0.0074和0.0004)。
hsa-miR-142-5p与Cyr61的相对表达量之间存在负相关性,相关系数为-0.9926(P=0.007)。
结论
1.Cyr61在胃癌细胞系BGC823,MGC803和SGC7901中高表达,在AGS中低表达;在新鲜胃癌和癌旁组织中的mRNA表达水平均高于正常胃黏膜组织。Cyr61定位于胃癌细胞的胞浆,在青年人胃癌和食管胃交界腺癌中均呈高表达。
2.青年人胃癌中,Cyr61在肠型胃癌比弥漫型胃癌表达高,其表达与临床分期呈正相关,临床分期越晚,Cyr61表达越高;Cyr61的表达与β-catenin、MMP9和HER2的表达具有相关性;单因素分析,β-catenin、MMP9阳性、PTEN阴性的患者预后比β-catenin、MMP9阴性、PTEN阳性的患者预后差。多因素分析,MMP9、CD44高表达的患者比低表达的患者预后差。
3.食管胃交界腺癌中,Cyr61的表达与临床分期呈正相关,临床分期越晚,Cyr61表达越高;Cyr61阳性表达的患者预后差。单因素分析,MMP9阳性的患者的生存率比MMP9阴性患者的生存率低。多因素分析,MMP9、Cyr61高表达的患者比低表达的患者预后差。
4.青年人胃癌的特点是弥漫型胃癌所占比例高,女性更易患弥漫型胃癌,且预后更差。单因素分析影响青年人胃癌预后的因素有性别,嗜酸性粒细胞浸润,脉管癌栓,肿瘤浸润深度,肿瘤最大径,淋巴结转移,远处转移和临床分期;多因素COX风险比例模型中,影响因素有肿瘤浸润深度,淋巴结转移和远处转移。
5.食管胃交界腺癌的特点是男性高发,组织学分型以管状腺癌最常见。单因素分析影响食管胃交界腺癌预后的因素有肿瘤浸润深度,淋巴结转移和临床分期;多因素COX风险比例模型中,肿瘤浸润深度和淋巴结转移成为入选因素。
Objective
Gastric Cancer(GC)is the most common malignant tumor of the digestive tract.At present,few molecular markers are found for early diagnosis,targeted therapy and prognosis predicting.Seeking for new clinical relevant molecular markers becomes the hotspot of GC research.The previous work of our laboratory on the gene expression profiling of GC tissue revealed that gene Cysteine-rich 61(Cyr61/CCN1)was remarkably over expressed in tumor tissue,pericancerous normal appearing tissue and normal gastric mucosa,which may play an important role in GC.In order to prove the results from the gene profiling data,we analyzed the transcription level of Cyr61 in gastric cancer cell lines,fresh tissue and the protein level of Cyr61 in 306 paraffin-embedded samples of GC(128 cases from the young age group;191 cases from the oesophagogastric junction adenocarcinoma,AEG).In recent years,though the morbility and mortality are decreasing,the incidence of GC at the young age and AEG are increasing.Little is known about these two special types of GC.Therefore,we detected the expression of Cyr61 and a panel of other markers which may relate to Cyr61 in these two special types of GC,expecting to verify the significance of Cyr61 in the prognosis of GC.Besides,the expression of microRNA(has-miR-142-Sp)which may regulate Cyr61 was detected in 4 gastric cancer cell lines.
Materials and Methods
306 cases of GC were collected and reviewed from the archives of the department of pathology in PLA General Hospital during the time period from 1996 to 2005.Tissue array were made.Cases with follow-up information were evaluated to identify valuable prognostic factors.
Envision procedure of immunohistochemistry was applied to detect the expression of Cyr61,CD44,CDX2,β-catenin,PTEN,MMP9,HER1,and HER2 on the paraffin-embedded tissue array sections.The expression of Cyr61 at the mRNA level in GC cell lines and fresh tissue samples was detected by using the methods of RT-PCR and Real time PCR.The location of Cyr61 protein was confirmed by immunofluorescence.Fluorescence in situ hybridization(FISH)was used to detect the amplification of HER2 in GC.The transcription level of microRNA in GC cell lines was detected by Real-Time PCR.The results were analyzed statistically with Kaplan-Meier plots and COX proportional hazard model by applying SPSS 13.0 software.The statistical significance level was set at P＜0.05.
Results
The mean age of GC of the young was 35.6 years with the range from 19 to 40 years.The male to female ratio was 1.91:1.The mean age of the AEG was 59.7 years with the range from 28 to 83 years.The male to female ratio was 8.1:1.121 cases of the young and 54 cases of AEG were followed-up.The longest survival time of the young patient who died of the tumor was 127.2 months,the shortest was 3.4 months.The longest survival time of the AEG patient who died of the tumor was 86 months,the shortest was 3 months.
Statistic analysis showed that in the young GC patients,vascular tumor emboli,female,eosinophile infiltration,depth of tumor invasion,large tumor diameter,lymph node metastases,distant metastasis,and late clinical stage were indicators of worse prognosis.In AEG,the depth of tumor invasion,lymph node metastasis,and late clinical stage correlated with poor outcome.The COX analysis revealed that depth of tumor invasion(P=0.0076),lymph node metastasis(P=0.0368),and distant metastasis(P=0.0156)were significantly correlated to the prognosis of GC of the young;depth of tumor invasion (P=0.0458),and lymph node metastasis(P=0.0389)significantly correlated to the prognosis of the AEG.
The highest expression of Cyr61 at the mRNA level in 6 GC cell lines was in BGC823,and the lowest was in AGS.The expression level of Cyr61 in GC tissue was much higher than it in the normal tissue.Cyr61 protein was located in cytoplasm,and its expression in the young GC patients was correlated to the WHO classification,Lauren classification,clinical stage,depth of tumor invasion, lymph node metastases,and distant metastases.The expression of Cyr61 in the AEG patients was correlated to the depth of tumor invasion,clinical stage,and shorter survival time(P=0.0001).
The expression of Cyr61 was correlated toβ-catenin,MMP9,and HER2 in young GC patients(P value=0.0009,0.0000 and 0.0000,respectively).The high expression ofβ-catenin(P=0.0314),MMP9(P=0.0112),and the loss of PTEN expression(P=0.0216)were associated with lower survival rate of young GC patients.The expression of Cyr61 was correlated to MMP9 in AEG patients (P=0.0368).The expression of MMP9(P=0.0451)was associated with lower survival rate in AEG patients.The COX analysis revealed that the high expression of MMP9 and CD44 in young GC patients and the high expression of MMP9 and Cyr61 in AEG patients were associated with lower survival rate.
There was a negative correlation between the expression of hsa-miR-142-5p and Cyr61(P=0.0007).
Conclusions
1.Cyr61 is highly expressed in GC cell lines BGC823,MGC303,and SGC7901, lowly expressed in AGS.Cyr61 mRNA expressed in GC tissue and pericancerous normal appearing tissue at a high level,which was consistent with the results of gene expression profiling.Cyr61 protein was located at the cytoplasm of GC tumor cell and was highly expressed both in young GC patients and AEG patients.
2.The expression of Cyr61 protein was more common in the intestinal type than in the diffuse type of young GC patients,and the expression had positive correlation with clinical stage,β-catenin,MMP9,and HER2.The expression ofβ-catenin,MMP9,and loss of PTEN was correlated to the low survival rate in young GC patient.The high expression of MMP9 and CD44 in young GC patients was associated with lower survival rate.
3.The expression of Cyr61 protein in AEG patients showed positive correlation to the clinical stage,and was associated with lower survival rate.MMP9 had low survival rate.The COX analysis revealed that the high expression of MMP9 and Cyr61 in AEG patients was associated with lower survival rate.
4.The main histological type of GC in young patients is diffuse type,with female predominance and worse prognosis.Vescular tumor emboli,female, eosinophile infiltration,deep tumor invasion,large tumor diameter,lymph node metastases,distant metastasis,and late clinical stage were indicators of worse prognosis.
5.The characteristics of AEG were male predominance,and with the main histological type of tubular adenocarcinoma.Deep tumor invasion,lymph node metastasis,and late clinical stage were bad indicators for prognosis.
引文
1.Wang XQ,Terry PD,Yan H.Review of salt consumption and stomach cancer risk:epidemiological and biological evidence.World J Gastroenterol.2009,May,15(18):2204-13.
2.Parkin DM,Bray F,Ferlay J,Pisani P.Estimating the world cancer burden:Globocan 2000.Int J Cancer.2001,94(2):153-156.
3.孙秀娣,牧人,周有尚等.中国胃癌死亡率20年变化情况分析及其发展趋势预测.中华肿瘤杂志.2004,26(1):4-9.
4.Mo FE,Muntean AG,Chen CC,et al.CYR61(CCNl)is essential for placental development and vascular integrity.Mol Cell Biol.2002,22(24):8709-8720.
5.Xie D,Yin D,Tong X,et al.Cyr61 is overexpressed in gliomas and involved in integrin-linked kinase-mediated Akt and beta-catenin-TCF/Lef signaling pathways.Cancer Res.2004,64:1987-1996.
6.Tong X,O'Kelly J,Xie D,et al.Cyr61 suppresses the growth of non-small-cell lung cancer cells via the beta-catenin-c-myc-p53 pathway.Oncogene.2004,23(28):4847-4855.
7.Maeta N,Osaki M,Shomori K,et al.CYR61 downregulation correlates with tumor progression by promoting MMP-7 expression in human gastric carcinoma.Oncology.2007;73(1-2):118-26.Epub 2008 Mar 13.
8.Lin MT,Chang CC,Chen ST,et al.Cyr61expression confers resistance to apoptosis in breast cancer MCF-7 cells by a mechanism of NF-κB-dependent XIAP up-regulation.J Biol Chem.2004,279(23):24015-24023.
9.Lin MT,Zuon CY,Chang CC,et al.Cyr61 induces gastric cancer cell motility/invasion via activation of the integrin/nuclear factor-kappaB/Cyclooxygennase-2 signaling pathway.Clin Cancer Res.2005,11(16):5809-5820.
10.Lin MT,Kuo IH,Chang CC,et al.Involvement of HIF-1-dependent PAI-1up-regulation in Cyr61/CCN1 induced gastric cancer cell invasion.J Biol Chem.2008,283(23):15807-15815.
11.Lin MT,Chang CC,Lin BR,et al.Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin alpha2betal.J Biol Chem.2007,282(47):34594-34604.
12.Hongo M.Review article:Barrett's oesophagus and carcinoma in Japan.Aliment.Pharmacol.Ther.2004,20(Suppl.8):50-54.
13.Fitzgerald RC.Review article:Barrett's oesophagus and associated adenocarcinoma-a UK perspective.Aliment.Pharmacol.Ther.2004,20(Suppl.8):45-49.
14.Parkin D.Max,Bray Freddie,Ferlay J,Pisani Paola.Global Cancer Statistics,2002.CA Cancer J Clin.2005,55:74-108.
15.Wong WM,Lai KC,Lam KF et al.Prevalence,clinical spectrum and health care utilization of gastro-oesophageal reflux disease in a Chinese population:a population-based study.Aliment.Pharmacol.Ther.2003,18:595-604.
16.张祥宏.食管胃交界腺癌诊断治疗的有关问题.中国肿瘤临床.2008,35(21):1201-1205.
17.张祥宏.食管胃交界腺癌——一个值得病理学家关注的问题.中华病理学杂志.2007,36(6):364-367.
18.尚晓滨,于振涛.食管胃交界部腺癌:概念与对策.中国癌症杂志.2008,
18(5):363-365.
19.Colin T.W,Julie Radeff-H,Rosalia M,et al.Thrombin receptor and RhoA mediate cell proliferation through integrins and cysteine-rich protein 61.FASEB.2008,22:4011-4021.
20.Stanley R,Lauri A,et al.Pathology and Genetics of Tumours of the Digestive System.IARC Press.2000,43-45.
21.Lai IR,Lee WJ,Chen CN,et al.Gastric cancer in the young.Hepatogastroenterology.1997,44(18):1641-1645.
22.Nakamura T,Yao T,Niho Y,et al.A clinicopathologic study in young patients with gastric carcinoma.J Surg Oncol.1999,71(4):211-219.
23.Eguchi T,Takahoshi Y,Yamagata M,et al.Gastric cancer in the young.Am Coll Surg.1999,188(1):22-26.
24.Lu C,Wang ZN,Sun Z,et al.Clinicopathologic features and prognosis of gastric cancer in young adults.Zhonghua Wai Ke Za Zhi.2008 Oct 1,46(19):1468-1471.
25.梁寒,郝希山,王家仓等.青年人胃癌.中华胃肠外科杂志.2001,4(2):85-87.
26.Theuer CP,de Virgilio C,Keese G,et al.Gastric adenocarcinoma in patients 40 years of age or younger.Am J Surg.1996,172(5):473-476.
27.Theuer CP,Kurosaki T,Taylor TH,et al.Unique Features of Gastric Carcinoma in the Young.Cancer.1998,83(1):25-33.
28.Fujimoto S,Takahashi M,Ohkubo H,et al.Comparative clinicopathologic features of early gastric cancer in young and older patients.Surgery.1994,115(4):516-520.
29.Walther C,Zilling T,Perfekt R,et al.Strongly increasing incidence of adenocarcinoma of the esophagus and gastric cardia.Lakartidningen.2004,101:180-183.
30.Kubo A,Corley DA.Marked regional variation in adenocarcinoma of the esophagus and the gastric cardia in the United States.Cancer 2002,95:2096-2102.
31.Tokunaga A,Nishi K,Matsukura N,et al.Estrogen and progesterone receptors in gastric cancer.Cancer.1986,57(7):1376-1379.
32.Harrison J,Jones J,Ellis I,et al.Oestrogen receptor D5 antibody is an independent negative prognostic factor in gastric cancer.Br J Surg.1991,78(3):334-336.
33.Roukos DH,Lorenz M,Encke A.Evidence of survival benefit of extended (D2)lymphadenectomy in western patients with gastric cancer based on a new concept:a prospective long-term follow-up study.Surgery.1998,123:573-578.
34.李华,路平,刘彩刚等.青年早期胃癌临床病理特征及预后因素探讨.中国普通外科杂志.2007,16(9):910-912.
35.杨维良,张新晨,张东伟等.35岁以下胃癌患者380例临床分析.中华胃肠 外科杂志.2004,7(5):357-359.
36.W.S.Lee et al.Prognostic significance of Lauren classification in curatively resected gastric cancer.2003 ASCO Annual Meeting.
37.Calli Demirkan et al.Correlation of histological classifications of gastric carcinomas with location and prognosis.Gastroenterol Clin Biol.2002 Jun-Jul,26(6-7):610-615.
38.Nielsen HJ,Hansen U,Christensen IJ,et al.Independent prognosis value of eosinophil and mast cell infiltration in colorectal cancer tissue.J Pathol.1999 Dec,189(4):487-495.
39.Kapp DS,Livolsi V.Intense eosinophilic stromal infiltrate in carcinoma of the uterine cervix:clinicopathologic study of 14 cases.Gynecologic oncologic 1983,16:19-30.
40.Lowe D,Fletcher CDM.Tumor-associated tissue eosinophilia in the bladder.J Clin Pathol.1984,37:500-502.
41.Lowe D,Fletcher CDM.Eosinophilia in squamous cell carcinoma of the oral cavity,external genitalia and anus-clinical correlatins.Histopathology 1984,8:
627-632.
42.王良俊,钱文英.间质反应细胞对进展期胃癌生物学行为的影响.滨州医学院学报.2006;29(4):261-263.
43.Hansen S,Whg J,Giercksky KE,Tretli S.Esophageal and gastric carcinoma in Norway 1958-1992:incidence time trend variability according to morphological subtypes and organ subsites.Int J Cancer.1997,71:340-344.
44.Shinichi H,Takaki Y,et al.Is Adenocarcinoma of the Esophagogastric Junction Different between Japan and Western Countries?The Incidence and Clinicopathological Features at a Japanese High-Volume Cancer Center.World J Surg.2009,33:95-103.45.Zhang Xun,David I Watson and Glyn G Jamieson.Lymph node metastases of adenocarcinoma of the esophagus and esophagogastric junction.Chin Med J.2007,120(24):2268-2270.
46.Sz(?)kacs G,Annereau JP,Lababidi S,et al.Predicting drug sensitivity and resistance:profiling ABC transporter genes in cancer cells.Cancer cell.2004,6:129-137.
47.Ohira M,Oba S,Nakamura Y,et al.Expression profiling using a tumor-specific cDNA microarray predicts the prognosis of intermediate risk neuroblastomas.Cancer Cell.2005 Apr,7(4):337-350.
48.Kim B,Bang S,Lee S,et al.Expression profiling and subtype-specific expression of stomach cancer.Cancer Res.2003,63:8248-8255.
49.Couzin J.Breakthrough of the year:Small RNAs make big splash.Science 2002,298:2296-2297.
50.Patricia S Steeg.Tumor metastasis:mechanistic insights and clinical challenges.Nature Medicine.2006,12(8):895-904.
51.张明基,应敏刚．WNT信号传导通路与胃癌．中国肿瘤．2007,16(11)：920-923．
52.S(?)rgio N,Jos(?)Carlos M,Carlos L,et al.Patterns of β-catenin expression in gastric carcinoma:clinicopathological relevance and mutation analysis.International Journal of Surgical Pathology.2003,11(1):1-9.
53.Gordana K,Sandra S.Matrix metalloproteinases in the process of invasion and metastasis of breast cancer.Arch Oncol.2006,14(3-4):136-40.
54.Egeblad,M,et al.New functions for the matrix metalloproteinases in cancer progression.Nat.Rev.Cancer.2002,2:161-174.
55.Kerkela E.et al.Matrix metalloproteinases in tumor progression:focus on basal and squamous cell skin cancer.Exp Dermatol.2003,12:109-125.
56.Vincenti,M.P.et al.Transcriptional regulation of collagenase (MMP-1,MMP-13) genes in arthritis:integration of complex signaling pathways for the recruitment of gene-specific transcription factors.Arthritis Res.2002,4:157-164.
57.Tzu-Wei T,Wei-Hung Y,Yuh-Tzy L,et al.Cyr61 increases migration and MMP-13 expression via avb3 integrin,FAK,ERK and AP-1-dependent pathway in human chondrosarcoma cells.Carcinogenesis.2009,30(2):258-268.
58.Chandrasekar B,Mummidi S,Mahimainathan L,et al.Interleukin-18-induced human coronary artery smooth muscle cell migration is dependent on NF-kappaBand AP-1-mediated matrix metalloproteinase-9 expression and is inhibited by atorvastatin.J Biol Chem.2006 Jun 2,281(22):15099-109.Epub 2006 Mar 22.
59.Chen CC,Mo FE,Lau LF,et al.The angiogenic factor Cyr61 activates a genetic program for wound healing in human skin fibroblasts.J Biol Chem.2001Dec 14,276(50):47329-37.Epub 2001 Oct 2.
60.左玉如,汪家敏.MMP-9和 uPA 在胃癌组织中的表达及相关性研究.中国血液流变学杂志.2003,13(04):352-354.
61.Schuell B,Gruenberger T,Scheithauer Wet al.HER 2/neu protein expression in colorectal cancer.BMC Cancer.2006,8(6):123.
62.Eltze E,Wu lfing C,Von Struensee D,et al.Cox-2 and Her2/neu co-expression in invasive bladder cancer.Int J Oncol.2005,26(6):1525-1531.
63.McKenzie S J,DeSombre KA,Bast BS et al.Serum levels of HER-2 neu (C-erbB-2)correlate with overexpression of p185neu in human ovarian cancer.Cancer.1993,71:3942-3946.
64.Hetzel D J,Wilson TO,Keeney GL,et al.HER-2/neu expression:a major prognostic factor in endometrial cancer.Gynecol Oncol.1992,47:179-185.
65.钱露,郭宁.原癌基因 HER2转录调控的研究进展.中国肿瘤生物治疗杂志.2005,12(3):225-228.
66.Luciano Vellon,Javier A.Menendez and Ruth Lupu.The CYR61-dependent up-regulation of α vβ3 integrin expression is a novel molecular response to Fatty Acid Synthase(FAS)inhibition in breast cancer cells.Proc Amer Assoc Cancer Res.2006,47.
67.Ambros V.,B.Bartel,D.P.Bartel,et al.A uniform system for microRNA annotation.RNA.2003,9:277-279.
68.Baehrecke,E.H.miRNAs:micro managers of programmed cell death.Curr.Biol.2003,13:473-475.
69.Bartel,D.P.MicroRNAs:genomics,biogenesis,mechanism,and function. Cell.2004,116:281-297.
70.Chan,J.A.,A.M.Krichevsky,and K.S.Kosik.MicroRNA-21 is an Anti-apoptotic factor in human glioblastoma cells.Cancer Res.2005,65:6029-6033.
71.Cimmino A.,G.A.Calin,M.Fabbri,et al.miR-15 and miR-16 induce apoptosis by targeting BCL2.Proc.Natl.Acad.Sci.2005,102:13944-13949.
72.Esquela-Kerscher.A.,and F.J.Slack.Oncomirs—microRNAs with a role in cancer.Nat.Rev.Cancer.2006,6:259-269.
73.Kluiver J.,S.Poppema,D.de Jong,et al.BIC and miR-155 are highly expressed in Hodgkin,primary mediastinal and diffuse large B cell lymphomas.J.Pathol.2005,207:243-249.
74.Brennecke J,Stark A,Russell R B,et al.Principles of microRNA-target recognition.PLoS Biol.2005,3:e85.
75.Enright A J,John B,Gaul U,et al.MicroRNA targets in Drosophila.Genome Biol.2003,5:R1.
76.Smith T F and Waterman M S.Identification of common molecular subsequences.J Mol Biol.1981,147:195-197.
77.Kiriakidou M,Nelson P T,Kouranov A,et al.A combined computational-experimental approach predicts human microRNA targets.Genes Dev,2004,18:1165-1178.
1.Parkin DM,Bray F,Ferlay J,Pisani P.Estimating the world cancer burden:Globocan 2000.Int J Cancer.2001,94(2):153-156.
2.Parkin DM,Pisani P,Ferlay J.Estimates of the worldwide incidence of 25major cancers in 1990.Int J Cancer.1999,80(6):827-841.
3.黄海力,王孟薇.肿瘤浸润转移分子机制的研究进展.生物技术通讯.2006,17(1):84-87.
4.Lau LF,Nathans D.Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells.EMBO J.1985,4(12):3145-3151.
5.Wong W,Kireeva M L,Kolesnikova T V,el al.Cyr61,product of a growth factor-inducible immediate-early gene,regulates chondrogenesis in mouse limb bud mesenchymal cells.Dev Biol.1997,192(2):492-508.
6.Mo FE,Muntean AG,Chen CC,et al.CYR61(CCN1)is essential for placental development and vascular integrity.Mol Cell Biol.2002,22(24):8709-8720.
7.Lechner A,Schutze N,Siggelkow H,et al.The immediate early gene product hCYR61 localizes to the secretory pathway in human osteoblasts.Bone.2000,27(1):53-60.
8.Hadjiargyrou M,Ahrens W & Rubin CT.Temporal expression of the chondrogenic and angiogenic growth factor CYR61 during fracture repair.J Bone Miner Res.2000,15(6):1014-1023.
9.Lienau J,Schell H,Epari DR,et al.CYR61(CCN1)protein expression during fracture healing in an ovine tibial model and its relation to the mechanical fixation stability.J Orthop Res.2005,24(2):254-262.
10.Ginsberg MH,Partridge A,Shattil SJ.Integrin regulation.Curr Opin Cell Biol.2005,17(5):509-516.
11.Kolesnikova TV,Lau LF.Human CYR61-mediated enhancement of bFGF -induced DNA synthesis in human umbilical vein endothelial cells.Oncogene.1998,16(6):747-754.
12.Schober JM,Lau LF,Ugarova TP,et al.Identification of a novel integrin alphaMbeta2 binding site in CCNI (CYR61),a matricellular protein expressed in healing wounds and atherosclerotic lesions.J Biol Chem.2003,278(28):25808-25815.
13.Chien W,Kumagai T,Miller CW,et al.Cyr61 suppresses growth of human endometrial cancer cells.J Biol Chem.2004,279(51):53087-53096.
14.Tong X,O'Kelly J,Xie D,et al.Cyr61 suppresses the growth of non-small-cell lung cancer cells via the beta-catenin-c-myc-p53 pathway.Oncogene.2004,23(28):4847-4855.
15.Tsai MS,Hornby AE,Lakins J,et al.Expression and function of CYR61,an angiogenic factor,in breast cancer cell lines and tumor biopsies.Cancer Res,2000,60(20):5603-5607.
16.Lin MT,Chang CC,Chen ST,et al.Cyr61 expression confers resistance to apoptosis in breast cancerMCF-7 cells by a mechanism of NF-KB-dependent XIAP up-regulation.J Biol Chem.2004,279(23):24015-23.
17.Xie D,Yin D,Tong X,et al.Cyr61 is overexpressed in gliomas and involved in integrin-linked kinase-mediated Akt and beta-catenin-TCF/Lef signaling pathways.Cancer Res.2004,64(6):1987-96.
18.Menendez JA,Mehmi I,Griggs DW,et al.The angiogenic factor CYR61 in breast cancer:molecular pathology and therapeutic perspectives.Endocr Relat Cancer.2003,10(2):141-52.
19.Tsai MS,Bogart DF,Li P,et al.Expression and regulation of Cyr61in human breast cancer cell lines.Oncogene.2002,21(6):964-73.
20.Hong Y,Ho K S,Eu K W,et al.A susceptibility gene set for early onset colorectal cancer that integrates diverse signaling pathways:implication for tumorigenesis.Clin Cancer Res.2007,13(4):1107-1114.
21.Babic AM,Kireeva ML,et al.Cyr61,a product of a growth factor-inducible immediate early gene,promotes angiogenesis and tumor growth.Proc.Natl.Acad.Sci.U.S.A.1998,95(11):6355-6360.
22.Lin MT,Zuon CY,Chang CC,et al.Cyr61 induces gastric cancer cell motility/invasion via activation of the integrin/nuclear factor-kappaB/Cyclooxyge-nase-2 signaling pathway.Clin Cancer Res.2005,11(16):5809-5820.
23.Lin MT,Chang CC,Lin BR,et al.Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin alpha2betal.J Biol Chem.2007,282(47):34594-34604.
24.Gately S,Li WW.Multiple roles of COX-2 in tumor angiogenesis:a target for antiangiogenic therapy.Semin Oncol.2004,31 (2 Suppl 7):2-11.
25.Uefuji K,Ichikura T,Mochizuki H.Cyclooxygenase-2 expression is related to prostaglandin biosynthesis and angiogenesis in human gastric cancer.Clin Cancer Res.2000,6(1):135-138.
26.Lin MT,Kuo IH,Chang CC,et al.Involvement of HIF-1-dependent PAI-1 up-regulation in Cyr61/CCN1 induced gastric cancer cell invasion.J Biol Chem.2008,283(23):15807-15.
27.Yoo,C.H.,Noh,S.H.,Shin,D.W.,et al.Recurrence following curative resection for gastric carcinoma.Br.J.Surg.2000,87(2):236-242.
28.Lin BR,Chang CC,Chen LR,et al.Cysteine-Rich 61(CCN1) enhances chemotactic migration,transendothelial cell migration,and intravasation by concomitantly up-regulating chemokine receptor 1 and 2.Mol Cancer Res.2007,5(11):1111-23.
2.Parkin DM,Bray F,Ferlay J,Pisani P.Estimating the world cancer burden:Globocan 2000.Int J Cancer.2001,94(2):153-156.
3.孙秀娣,牧人,周有尚等.中国胃癌死亡率20年变化情况分析及其发展趋势预测.中华肿瘤杂志.2004,26(1):4-9.
4.Mo FE,Muntean AG,Chen CC,et al.CYR61(CCNl)is essential for placental development and vascular integrity.Mol Cell Biol.2002,22(24):8709-8720.
5.Xie D,Yin D,Tong X,et al.Cyr61 is overexpressed in gliomas and involved in integrin-linked kinase-mediated Akt and beta-catenin-TCF/Lef signaling pathways.Cancer Res.2004,64:1987-1996.
6.Tong X,O'Kelly J,Xie D,et al.Cyr61 suppresses the growth of non-small-cell lung cancer cells via the beta-catenin-c-myc-p53 pathway.Oncogene.2004,23(28):4847-4855.
7.Maeta N,Osaki M,Shomori K,et al.CYR61 downregulation correlates with tumor progression by promoting MMP-7 expression in human gastric carcinoma.Oncology.2007;73(1-2):118-26.Epub 2008 Mar 13.
8.Lin MT,Chang CC,Chen ST,et al.Cyr61expression confers resistance to apoptosis in breast cancer MCF-7 cells by a mechanism of NF-κB-dependent XIAP up-regulation.J Biol Chem.2004,279(23):24015-24023.
9.Lin MT,Zuon CY,Chang CC,et al.Cyr61 induces gastric cancer cell motility/invasion via activation of the integrin/nuclear factor-kappaB/Cyclooxygennase-2 signaling pathway.Clin Cancer Res.2005,11(16):5809-5820.
10.Lin MT,Kuo IH,Chang CC,et al.Involvement of HIF-1-dependent PAI-1up-regulation in Cyr61/CCN1 induced gastric cancer cell invasion.J Biol Chem.2008,283(23):15807-15815.
11.Lin MT,Chang CC,Lin BR,et al.Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin alpha2betal.J Biol Chem.2007,282(47):34594-34604.
12.Hongo M.Review article:Barrett's oesophagus and carcinoma in Japan.Aliment.Pharmacol.Ther.2004,20(Suppl.8):50-54.
13.Fitzgerald RC.Review article:Barrett's oesophagus and associated adenocarcinoma-a UK perspective.Aliment.Pharmacol.Ther.2004,20(Suppl.8):45-49.
14.Parkin D.Max,Bray Freddie,Ferlay J,Pisani Paola.Global Cancer Statistics,2002.CA Cancer J Clin.2005,55:74-108.
15.Wong WM,Lai KC,Lam KF et al.Prevalence,clinical spectrum and health care utilization of gastro-oesophageal reflux disease in a Chinese population:a population-based study.Aliment.Pharmacol.Ther.2003,18:595-604.
16.张祥宏.食管胃交界腺癌诊断治疗的有关问题.中国肿瘤临床.2008,35(21):1201-1205.
17.张祥宏.食管胃交界腺癌——一个值得病理学家关注的问题.中华病理学杂志.2007,36(6):364-367.
18.尚晓滨,于振涛.食管胃交界部腺癌:概念与对策.中国癌症杂志.2008,
18(5):363-365.
19.Colin T.W,Julie Radeff-H,Rosalia M,et al.Thrombin receptor and RhoA mediate cell proliferation through integrins and cysteine-rich protein 61.FASEB.2008,22:4011-4021.
20.Stanley R,Lauri A,et al.Pathology and Genetics of Tumours of the Digestive System.IARC Press.2000,43-45.
21.Lai IR,Lee WJ,Chen CN,et al.Gastric cancer in the young.Hepatogastroenterology.1997,44(18):1641-1645.
22.Nakamura T,Yao T,Niho Y,et al.A clinicopathologic study in young patients with gastric carcinoma.J Surg Oncol.1999,71(4):211-219.
23.Eguchi T,Takahoshi Y,Yamagata M,et al.Gastric cancer in the young.Am Coll Surg.1999,188(1):22-26.
24.Lu C,Wang ZN,Sun Z,et al.Clinicopathologic features and prognosis of gastric cancer in young adults.Zhonghua Wai Ke Za Zhi.2008 Oct 1,46(19):1468-1471.
25.梁寒,郝希山,王家仓等.青年人胃癌.中华胃肠外科杂志.2001,4(2):85-87.
26.Theuer CP,de Virgilio C,Keese G,et al.Gastric adenocarcinoma in patients 40 years of age or younger.Am J Surg.1996,172(5):473-476.
27.Theuer CP,Kurosaki T,Taylor TH,et al.Unique Features of Gastric Carcinoma in the Young.Cancer.1998,83(1):25-33.
28.Fujimoto S,Takahashi M,Ohkubo H,et al.Comparative clinicopathologic features of early gastric cancer in young and older patients.Surgery.1994,115(4):516-520.
29.Walther C,Zilling T,Perfekt R,et al.Strongly increasing incidence of adenocarcinoma of the esophagus and gastric cardia.Lakartidningen.2004,101:180-183.
30.Kubo A,Corley DA.Marked regional variation in adenocarcinoma of the esophagus and the gastric cardia in the United States.Cancer 2002,95:2096-2102.
31.Tokunaga A,Nishi K,Matsukura N,et al.Estrogen and progesterone receptors in gastric cancer.Cancer.1986,57(7):1376-1379.
32.Harrison J,Jones J,Ellis I,et al.Oestrogen receptor D5 antibody is an independent negative prognostic factor in gastric cancer.Br J Surg.1991,78(3):334-336.
33.Roukos DH,Lorenz M,Encke A.Evidence of survival benefit of extended (D2)lymphadenectomy in western patients with gastric cancer based on a new concept:a prospective long-term follow-up study.Surgery.1998,123:573-578.
34.李华,路平,刘彩刚等.青年早期胃癌临床病理特征及预后因素探讨.中国普通外科杂志.2007,16(9):910-912.
35.杨维良,张新晨,张东伟等.35岁以下胃癌患者380例临床分析.中华胃肠 外科杂志.2004,7(5):357-359.
36.W.S.Lee et al.Prognostic significance of Lauren classification in curatively resected gastric cancer.2003 ASCO Annual Meeting.
37.Calli Demirkan et al.Correlation of histological classifications of gastric carcinomas with location and prognosis.Gastroenterol Clin Biol.2002 Jun-Jul,26(6-7):610-615.
38.Nielsen HJ,Hansen U,Christensen IJ,et al.Independent prognosis value of eosinophil and mast cell infiltration in colorectal cancer tissue.J Pathol.1999 Dec,189(4):487-495.
39.Kapp DS,Livolsi V.Intense eosinophilic stromal infiltrate in carcinoma of the uterine cervix:clinicopathologic study of 14 cases.Gynecologic oncologic 1983,16:19-30.
40.Lowe D,Fletcher CDM.Tumor-associated tissue eosinophilia in the bladder.J Clin Pathol.1984,37:500-502.
41.Lowe D,Fletcher CDM.Eosinophilia in squamous cell carcinoma of the oral cavity,external genitalia and anus-clinical correlatins.Histopathology 1984,8:
627-632.
42.王良俊,钱文英.间质反应细胞对进展期胃癌生物学行为的影响.滨州医学院学报.2006;29(4):261-263.
43.Hansen S,Whg J,Giercksky KE,Tretli S.Esophageal and gastric carcinoma in Norway 1958-1992:incidence time trend variability according to morphological subtypes and organ subsites.Int J Cancer.1997,71:340-344.
44.Shinichi H,Takaki Y,et al.Is Adenocarcinoma of the Esophagogastric Junction Different between Japan and Western Countries?The Incidence and Clinicopathological Features at a Japanese High-Volume Cancer Center.World J Surg.2009,33:95-103.45.Zhang Xun,David I Watson and Glyn G Jamieson.Lymph node metastases of adenocarcinoma of the esophagus and esophagogastric junction.Chin Med J.2007,120(24):2268-2270.
46.Sz(?)kacs G,Annereau JP,Lababidi S,et al.Predicting drug sensitivity and resistance:profiling ABC transporter genes in cancer cells.Cancer cell.2004,6:129-137.
47.Ohira M,Oba S,Nakamura Y,et al.Expression profiling using a tumor-specific cDNA microarray predicts the prognosis of intermediate risk neuroblastomas.Cancer Cell.2005 Apr,7(4):337-350.
48.Kim B,Bang S,Lee S,et al.Expression profiling and subtype-specific expression of stomach cancer.Cancer Res.2003,63:8248-8255.
49.Couzin J.Breakthrough of the year:Small RNAs make big splash.Science 2002,298:2296-2297.
50.Patricia S Steeg.Tumor metastasis:mechanistic insights and clinical challenges.Nature Medicine.2006,12(8):895-904.
51.张明基,应敏刚．WNT信号传导通路与胃癌．中国肿瘤．2007,16(11)：920-923．
52.S(?)rgio N,Jos(?)Carlos M,Carlos L,et al.Patterns of β-catenin expression in gastric carcinoma:clinicopathological relevance and mutation analysis.International Journal of Surgical Pathology.2003,11(1):1-9.
53.Gordana K,Sandra S.Matrix metalloproteinases in the process of invasion and metastasis of breast cancer.Arch Oncol.2006,14(3-4):136-40.
54.Egeblad,M,et al.New functions for the matrix metalloproteinases in cancer progression.Nat.Rev.Cancer.2002,2:161-174.
55.Kerkela E.et al.Matrix metalloproteinases in tumor progression:focus on basal and squamous cell skin cancer.Exp Dermatol.2003,12:109-125.
56.Vincenti,M.P.et al.Transcriptional regulation of collagenase (MMP-1,MMP-13) genes in arthritis:integration of complex signaling pathways for the recruitment of gene-specific transcription factors.Arthritis Res.2002,4:157-164.
57.Tzu-Wei T,Wei-Hung Y,Yuh-Tzy L,et al.Cyr61 increases migration and MMP-13 expression via avb3 integrin,FAK,ERK and AP-1-dependent pathway in human chondrosarcoma cells.Carcinogenesis.2009,30(2):258-268.
58.Chandrasekar B,Mummidi S,Mahimainathan L,et al.Interleukin-18-induced human coronary artery smooth muscle cell migration is dependent on NF-kappaBand AP-1-mediated matrix metalloproteinase-9 expression and is inhibited by atorvastatin.J Biol Chem.2006 Jun 2,281(22):15099-109.Epub 2006 Mar 22.
59.Chen CC,Mo FE,Lau LF,et al.The angiogenic factor Cyr61 activates a genetic program for wound healing in human skin fibroblasts.J Biol Chem.2001Dec 14,276(50):47329-37.Epub 2001 Oct 2.
60.左玉如,汪家敏.MMP-9和 uPA 在胃癌组织中的表达及相关性研究.中国血液流变学杂志.2003,13(04):352-354.
61.Schuell B,Gruenberger T,Scheithauer Wet al.HER 2/neu protein expression in colorectal cancer.BMC Cancer.2006,8(6):123.
62.Eltze E,Wu lfing C,Von Struensee D,et al.Cox-2 and Her2/neu co-expression in invasive bladder cancer.Int J Oncol.2005,26(6):1525-1531.
63.McKenzie S J,DeSombre KA,Bast BS et al.Serum levels of HER-2 neu (C-erbB-2)correlate with overexpression of p185neu in human ovarian cancer.Cancer.1993,71:3942-3946.
64.Hetzel D J,Wilson TO,Keeney GL,et al.HER-2/neu expression:a major prognostic factor in endometrial cancer.Gynecol Oncol.1992,47:179-185.
65.钱露,郭宁.原癌基因 HER2转录调控的研究进展.中国肿瘤生物治疗杂志.2005,12(3):225-228.
66.Luciano Vellon,Javier A.Menendez and Ruth Lupu.The CYR61-dependent up-regulation of α vβ3 integrin expression is a novel molecular response to Fatty Acid Synthase(FAS)inhibition in breast cancer cells.Proc Amer Assoc Cancer Res.2006,47.
67.Ambros V.,B.Bartel,D.P.Bartel,et al.A uniform system for microRNA annotation.RNA.2003,9:277-279.
68.Baehrecke,E.H.miRNAs:micro managers of programmed cell death.Curr.Biol.2003,13:473-475.
69.Bartel,D.P.MicroRNAs:genomics,biogenesis,mechanism,and function. Cell.2004,116:281-297.
70.Chan,J.A.,A.M.Krichevsky,and K.S.Kosik.MicroRNA-21 is an Anti-apoptotic factor in human glioblastoma cells.Cancer Res.2005,65:6029-6033.
71.Cimmino A.,G.A.Calin,M.Fabbri,et al.miR-15 and miR-16 induce apoptosis by targeting BCL2.Proc.Natl.Acad.Sci.2005,102:13944-13949.
72.Esquela-Kerscher.A.,and F.J.Slack.Oncomirs—microRNAs with a role in cancer.Nat.Rev.Cancer.2006,6:259-269.
73.Kluiver J.,S.Poppema,D.de Jong,et al.BIC and miR-155 are highly expressed in Hodgkin,primary mediastinal and diffuse large B cell lymphomas.J.Pathol.2005,207:243-249.
74.Brennecke J,Stark A,Russell R B,et al.Principles of microRNA-target recognition.PLoS Biol.2005,3:e85.
75.Enright A J,John B,Gaul U,et al.MicroRNA targets in Drosophila.Genome Biol.2003,5:R1.
76.Smith T F and Waterman M S.Identification of common molecular subsequences.J Mol Biol.1981,147:195-197.
77.Kiriakidou M,Nelson P T,Kouranov A,et al.A combined computational-experimental approach predicts human microRNA targets.Genes Dev,2004,18:1165-1178.
1.Parkin DM,Bray F,Ferlay J,Pisani P.Estimating the world cancer burden:Globocan 2000.Int J Cancer.2001,94(2):153-156.
2.Parkin DM,Pisani P,Ferlay J.Estimates of the worldwide incidence of 25major cancers in 1990.Int J Cancer.1999,80(6):827-841.
3.黄海力,王孟薇.肿瘤浸润转移分子机制的研究进展.生物技术通讯.2006,17(1):84-87.
4.Lau LF,Nathans D.Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells.EMBO J.1985,4(12):3145-3151.
5.Wong W,Kireeva M L,Kolesnikova T V,el al.Cyr61,product of a growth factor-inducible immediate-early gene,regulates chondrogenesis in mouse limb bud mesenchymal cells.Dev Biol.1997,192(2):492-508.
6.Mo FE,Muntean AG,Chen CC,et al.CYR61(CCN1)is essential for placental development and vascular integrity.Mol Cell Biol.2002,22(24):8709-8720.
7.Lechner A,Schutze N,Siggelkow H,et al.The immediate early gene product hCYR61 localizes to the secretory pathway in human osteoblasts.Bone.2000,27(1):53-60.
8.Hadjiargyrou M,Ahrens W & Rubin CT.Temporal expression of the chondrogenic and angiogenic growth factor CYR61 during fracture repair.J Bone Miner Res.2000,15(6):1014-1023.
9.Lienau J,Schell H,Epari DR,et al.CYR61(CCN1)protein expression during fracture healing in an ovine tibial model and its relation to the mechanical fixation stability.J Orthop Res.2005,24(2):254-262.
10.Ginsberg MH,Partridge A,Shattil SJ.Integrin regulation.Curr Opin Cell Biol.2005,17(5):509-516.
11.Kolesnikova TV,Lau LF.Human CYR61-mediated enhancement of bFGF -induced DNA synthesis in human umbilical vein endothelial cells.Oncogene.1998,16(6):747-754.
12.Schober JM,Lau LF,Ugarova TP,et al.Identification of a novel integrin alphaMbeta2 binding site in CCNI (CYR61),a matricellular protein expressed in healing wounds and atherosclerotic lesions.J Biol Chem.2003,278(28):25808-25815.
13.Chien W,Kumagai T,Miller CW,et al.Cyr61 suppresses growth of human endometrial cancer cells.J Biol Chem.2004,279(51):53087-53096.
14.Tong X,O'Kelly J,Xie D,et al.Cyr61 suppresses the growth of non-small-cell lung cancer cells via the beta-catenin-c-myc-p53 pathway.Oncogene.2004,23(28):4847-4855.
15.Tsai MS,Hornby AE,Lakins J,et al.Expression and function of CYR61,an angiogenic factor,in breast cancer cell lines and tumor biopsies.Cancer Res,2000,60(20):5603-5607.
16.Lin MT,Chang CC,Chen ST,et al.Cyr61 expression confers resistance to apoptosis in breast cancerMCF-7 cells by a mechanism of NF-KB-dependent XIAP up-regulation.J Biol Chem.2004,279(23):24015-23.
17.Xie D,Yin D,Tong X,et al.Cyr61 is overexpressed in gliomas and involved in integrin-linked kinase-mediated Akt and beta-catenin-TCF/Lef signaling pathways.Cancer Res.2004,64(6):1987-96.
18.Menendez JA,Mehmi I,Griggs DW,et al.The angiogenic factor CYR61 in breast cancer:molecular pathology and therapeutic perspectives.Endocr Relat Cancer.2003,10(2):141-52.
19.Tsai MS,Bogart DF,Li P,et al.Expression and regulation of Cyr61in human breast cancer cell lines.Oncogene.2002,21(6):964-73.
20.Hong Y,Ho K S,Eu K W,et al.A susceptibility gene set for early onset colorectal cancer that integrates diverse signaling pathways:implication for tumorigenesis.Clin Cancer Res.2007,13(4):1107-1114.
21.Babic AM,Kireeva ML,et al.Cyr61,a product of a growth factor-inducible immediate early gene,promotes angiogenesis and tumor growth.Proc.Natl.Acad.Sci.U.S.A.1998,95(11):6355-6360.
22.Lin MT,Zuon CY,Chang CC,et al.Cyr61 induces gastric cancer cell motility/invasion via activation of the integrin/nuclear factor-kappaB/Cyclooxyge-nase-2 signaling pathway.Clin Cancer Res.2005,11(16):5809-5820.
23.Lin MT,Chang CC,Lin BR,et al.Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin alpha2betal.J Biol Chem.2007,282(47):34594-34604.
24.Gately S,Li WW.Multiple roles of COX-2 in tumor angiogenesis:a target for antiangiogenic therapy.Semin Oncol.2004,31 (2 Suppl 7):2-11.
25.Uefuji K,Ichikura T,Mochizuki H.Cyclooxygenase-2 expression is related to prostaglandin biosynthesis and angiogenesis in human gastric cancer.Clin Cancer Res.2000,6(1):135-138.
26.Lin MT,Kuo IH,Chang CC,et al.Involvement of HIF-1-dependent PAI-1 up-regulation in Cyr61/CCN1 induced gastric cancer cell invasion.J Biol Chem.2008,283(23):15807-15.
27.Yoo,C.H.,Noh,S.H.,Shin,D.W.,et al.Recurrence following curative resection for gastric carcinoma.Br.J.Surg.2000,87(2):236-242.
28.Lin BR,Chang CC,Chen LR,et al.Cysteine-Rich 61(CCN1) enhances chemotactic migration,transendothelial cell migration,and intravasation by concomitantly up-regulating chemokine receptor 1 and 2.Mol Cancer Res.2007,5(11):1111-23.