Lentivirus-mediated RNA silencing of c-Met markedly suppresses peritoneal dissemination of gastric cancer in vitro and in vivo
Lentivirus-mediated RNA silencing of c-Met markedly suppresses peritoneal dissemination of gastric cancer in vitro and in vivo
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DOI：
10.1038/aps.2011.205
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发表时间：
2012-04-01
影响因子：
8.2
通讯作者：
Yang, Chang-qin
中科院分区：
文献类型：
--
作者：
Wang, Xiao-lei;Chen, Xi-mei;Yang, Chang-qin
目的：目的探讨胃癌腹水中游离癌细胞c-Met的表达及其与胃癌腹膜转移的关系。方法：从胃癌患者腹水中分离腹膜游离癌细胞（PFCCs）。免疫细胞化学法检测PFCCs中c-Met的表达。RT-PCR和Western blot检测胃癌细胞株SGC 7901中c-Met的表达，慢病毒介导的RNAi抑制c-Met的表达。MU法检测细胞增殖能力，侵袭实验检测细胞侵袭能力。在人腹膜间皮细胞（HPMCs）单层上观察了胃癌细胞（SGC 7901）在体外的粘附，在小鼠体内观察了胃癌细胞在腹膜上的粘附。结果：10例胃癌患者中有6例腹水中分离到PFCCs。6例胃癌患者中5例PFCCs表达c-Met。慢病毒介导的RNAi可显著抑制胃癌细胞株SGC 7901中c-Met mRNA和蛋白的表达，从而抑制胃癌细胞的增殖、侵袭和粘附。转染Lenti-miRNAc-Met的胃癌细胞la 3131整合素和E-cadherin的表达明显受到抑制。在胃癌腹膜转移模型中，腹腔注射Lenti-miRNAc-Met可明显抑制胃癌细胞株SGC 7901的肿瘤进展。结论：胃癌患者腹水中PFCCs表达c-Met。下调c-Met表达可明显抑制胃癌腹膜播散的多步骤过程，可能成为胃癌治疗的一个潜在靶点。
Aim: To investigate the expression of c-Met in peritoneal free cancer cells isolated from human gastric cancer ascites, and its relationship to peritoneal dissemination of gastric cancer. Methods: Peritoneal free cancer cells (PFCCs) were isolated from ascites specimens of gastric cancer patients. c-Met expression in PFCCs was detected with immunocytochemistry. In human gastric cancer cell line SGC7901, c-Met expression was detected using RT-PCR and Western blot, and was suppressed with lentivirus-mediated RNAi. The proliferation of SGC7901 cells was measured using MU assay, and the invasion ability was detected with invasion assay. The adhesion of SGC7901 cells to peritoneum was observed in human peritoneal mesothelial cells (HPMCs) monolayer in vitro and in mice in vivo. Results: PFCCs were isolated from ascites of 6 out of 10 gastric cancer patients. c-Met expression in PFCCs was detected in 5 of the 6 gastric cancer patients. In SGC7901 cells, Lentivirus-mediated RNAi significantly reduced both c-Met mRNA and protein expression, which resulted in suppressing the cell proliferation, invasion and adhesion to peritoneum. The expression of la3131 integrin and E-cadherin was significantly inhibited in SGC7901 cells transfected with Lenti-miRNAc-Met. In the peritoneal dissemination model of gastric cancer, intraperitoneal injection of Lenti-miRNAc-Met markedly suppressed the tumor progression of SGC7901 cells. Conclusion: c-Met is expressed in PFCCs from the ascites of gastric cancer patients. Down-regulation of c-Met expression markedly suppresses the multistep process of peritoneal dissemination, thus may be a potential target for the treatment of gastric cancer.