肿瘤相关成纤维细胞对胃癌细胞增殖和侵袭能力的影响及其机制研究

The effect of cancer-associated fibroblasts from gastric cancer on the invasiveness of HGC-27 cell

[目的]观察肿瘤相关成纤维细胞(CAFs)对胃癌HGC-27细胞侵袭能力的影响,并探讨其可能的作用机制。[方法]取胃癌患者癌组织及癌旁正常胃黏膜组织分别分离培养CAFs和正常成纤维细胞(normal fibroblasts,NFs),建立胃癌HGC-27细胞与CAFs或NFs共培养体系,CCK-8比色法检测细胞增殖,Transwell小室法检测细胞侵袭能力的改变,免疫印迹法检测细胞上皮间质转化(EMT)标志物[E-钙粘素(E-cadherin)蛋白、波形蛋白(Vimentin)和纤维连接蛋白(Fibronectin)蛋白]、MMP-9、STAT3、p-STAT3蛋白表达的改变。[结果]胃癌HGC-27细胞与CAFs共培养后其增殖和侵袭能力均较HGC-27细胞单独培养组或与NFs共培养组显著增强,且E-cadherin蛋白表达明显下调,同时Vimentin、Fibronectin蛋白的表达明显上调,MMP-9蛋白表达上调;STAT3总量未发生变化,p-STAT3表达上调。[结论]CAFs可促进胃癌细胞的侵袭能力,其机制可能是通过激活STAT3通路进而促进细胞EMT。

[Objective]To observe the effect of cancer-associated fibroblasts from gastric cancer on the invasiveness of HGC-27 cells,and explore the possible mechanisms.[Methods]Culturing CAFs and normal fibroblasts(NFs)isolated respectively from the tumor and adjacent normal gastric tissue.After establishing co-cultured system,the cell survival was determined by the CCK-8 method.The invasion of HGC-27 cells were tested by transwell assay.Western blotting was used for E-cadherin,Vimentin,Fibronectin,MMP-9,STAT3 and p-STAT3 protein analysis.[Results]Co-cultured with CAFs,the proliferation and invasiveness abilities of HGC-27 cells were significantly enhanced(P<0.05).The expression levels of MMP-9 and p-STAT3 were enhanced.Also the proteins levels of Vimentin and Fibronectin were increased,but the expression level of E-cadherin was decreased compared to gastric cancer cells group or NFs group.[Conclusion]CAFs can promote the proliferation and invasive abilities of HGC-27 cells,which is associated with promotion of the STAT3 signaling pathway,thus inducing epithelial-to-mesenchymal transition in gastric cancer cell line HGC-27 cells.