接头蛋白LNK抑制胃癌细胞增殖和侵袭能力的研究

The study on the mechanism of adaptor protein LNK inhibiting the proliferation and invasion of gastric cancer cells

为分析人接头蛋白LNK(hLNK)对胃癌发生发展的影响,克隆并包装含hLNK基因的重组慢病毒并感染人胃癌细胞株SGC-7901以建立稳定过表达hLNK的细胞系SGC-7901-hLNK(7901-hLNK);Western blot验证该稳转细胞系中hLNK的表达水平,平板克隆试验分析hLNK过表达对其增殖的影响;Transwell小室试验分析hLNK过表达对胃癌细胞侵袭能力的影响;并对细胞内信号分子p-Jak2、p-Stat3、p-p38、Jak2、Stat3、p38的活化水平进行分析。结果表明:过表达hLNK的人胃癌细胞株7901-hLNK得到成功构建,该细胞的增殖、侵袭能力较对照组明显被抑制,且细胞中p-Jak2、p-Stat3水平显著下调。这一研究提示,hLNK可能作为一种抑癌因子在胃癌中发挥作用,其主要通过下调Jak2-Stat3信号通路抑制癌细胞的增殖、侵袭。

To analyze the mechanism of human adaptor protein LNK(hLNK)in inhibiting the tumorigenesis and progression of gastric cancer cell,hLNK cDNA was amplified and subcloned to lentiviral vector pCDH to construct recombinant plasmid pCDH-hLNK,then the recombinant virus containing hLNK was packaged by transfecting 293 T cells with pCDH-hLNK and its packaging plasmids.The human gastric cancer cell line SGC-7901 was infected by recombinant virus and its control virus to establish cell line SGC-7901-hLNK(7901-hLNK)with the stable overexpression of hLNK.Then 7901-hLNK was tested by fluorescence microscopy and Western blot assay for the forced expresison of hLNK.Plate colony assay was used to analyze the proliferative capacity of 7901-hLNK,transwell chamber assay was performed to detect the metastasis ability of 7901-hLNK and the signaling pathway molecules Jak2,Stat3,p38 were analyzed by Western blot.The results indicated that 7901-hLNK with constant overexpression of hLNK was successfully constructed,which could strongly inhibit the proliferation and metastasis of 7901-hLNK with the decreased level of p-Jak2,p-Stat3.This study demonstrated that hLNK has strong inhibitory effect on the proliferation and metastasis of gastric cancer cell by down-regulating Jak2-Stat3 signaling pathway,indicating that hLNK works as a tumor suppressor in gastric cancer.