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目的了解表皮生长因子受体(epidermal growth factor receptor,EGFR)和整合素信号通路在胃癌细胞SGC7901中的交叉反应和对细胞侵袭增殖的影响。方法使用EGF和Fn刺激SGC7901细胞,免疫沉淀和蛋白质电泳检测ERK、FAK和p130cas总蛋白和FAK Y397、p130cas Y410和ERK总酪氨酸磷酸化的改变;使用改良Boyden小室法检测胃癌细胞侵袭力;MTT法检测细胞增殖的改变;使用RNA干扰降低FAK表达,观察FAK低表达对交叉反应和胃癌侵袭增殖的影响。结果 ERK、FAK和p130cas总蛋白在刺激前后无变化(P>0.05)。Fn刺激后,ERK总酪氨酸磷酸化增强了2.90倍(P<0.05),细胞侵袭力增强了2.36倍(P<0.05),24 h MTT值升高了1.68倍(P<0.05);EGF刺激后,FAKY397磷酸化增强了2.75倍(P<0.05),p130cas Y410磷酸化增强了4.33倍(P<0.05),细胞侵袭力增强了1.50倍(P<0.05),24 h MTT值分别升高了1.76倍(P<0.05)。转染FAK siRNA组细胞,FAK Y397磷酸化表达只有对照组的0.30倍(P<0.05);Fn刺激后,ERK总磷酸化表达只有对照组的0.66倍(P<0.05),细胞侵袭力只有对照组的0.37倍(P<0.05),24 h MTT值只有对照组的0.63倍(P<0.05);EGF刺激后,p130 Y410磷酸化只有对照组的0.49倍(P<0.05),细胞侵袭力只有对照组的0.48倍(P<0.05),24 h MTT值只有对照组0.77倍(P<0.05)。结论 EGFR和整合素信号在胃癌细胞中发生交叉反应,FAK是其中的关键信号分子,阻断FAK表达可以有效抑制两条信号通路引起的胃癌细胞侵袭和增殖。
Objective To investigate the effects of epidermal growth factor receptor (EGFR) and integrin signaling pathway on the cross-reactivity of SGC7901 cells and invasion and metastasis of gastric cancer cells. Methods SGC7901 cells were stimulated with EGF and Fn, and the total tyrosine phosphorylation of ERK, FAK and p130cas total proteins and the total tyrosine phosphorylation of FAK Y397, p130cas Y410 and ERK were detected by immunoprecipitation and protein electrophoresis. The invasiveness of gastric cancer cells was detected by modified Boyden chamber assay. MTT assay was used to detect the changes of cell proliferation; RNAi was used to reduce the expression of FAK; and the effect of low FAK expression on the cross-reactivity and gastric cancer invasion and proliferation was observed. Results The total protein of ERK, FAK and p130cas did not change before and after stimulation (P> 0.05). After Fn stimulation, total tyrosine phosphorylation of ERK increased by 2.90-fold (P <0.05), cell invasiveness increased by 2.36-fold (P <0.05) and MTT increased by 1.68-fold The phosphorylation of FAKY397 increased by 2.75 folds (P <0.05), the phosphorylation of p130cas Y410 increased by 4.33 folds (P <0.05), and the invasiveness of cells increased by 1.50 folds (P <0.05) 1.76 times (P <0.05). The expression of phosphorylated FAK Y397 in transfected FAK siRNA group was only 0.30 times that of the control group (P <0.05). After Fn stimulation, the total phosphorylation of ERK was only 0.66 times that of the control group (P <0.05) (P <0.05). The MTT value at 24 h was only 0.63 times that of the control group (P <0.05). The phosphorylation of p130 Y410 was only 0.49 times that of the control group (P <0.05) (P <0.05). The 24 h MTT value was only 0.77 times that of the control group (P <0.05). Conclusion EGFR and integrin signaling cross-react in gastric cancer cells. FAK is one of the key signaling molecules. Blocking FAK expression can effectively inhibit the invasion and proliferation of gastric cancer cells induced by the two signaling pathways.