Smad1在胃腺癌组织中的表达及对胃腺癌细胞迁移能力的影响研究

Study on expression of Smad1 in gastric adenocarcinoma and its effect on migration ability of gastric cancer cells

目的:研究Smad1在胃腺癌组织中的表达及对胃腺癌细胞迁移能力的影响。方法:提取收集的胃腺癌组织及对应的癌旁组织中的蛋白,Western blot检测Smad1的表达水平。以胃腺癌细胞HGC-27为研究对象,细胞转染过表达Smad1的载体(p-EGFP-C1/Smad1)和Smad1小干扰RNA(Smad1 siRNA),同时转染p-EGFP-C1和siRNA control为对照。细胞划痕实验检测细胞迁移能力。Western blot检测细胞中MMP-9、MMP-2、p-Akt、Akt的表达水平。Akt信号通路抑制剂LY294002(20μg/ml)作用于胃腺癌细胞HGC-27,MTT检测细胞增殖情况,细胞划痕实验检测细胞迁移能力。Western blot检测MMP-9、MMP-2、p-Akt、Akt蛋白表达水平。结果:胃腺癌组织中Smad1的水平明显低于癌旁组织(P<0.01)。p-EGFP-C1/Smad1组细胞存活率和迁移率均明显低于p-EGFP-C1组(P<0.01)。Smad1 siRNA组细胞存活率和迁移率均明显高于siRNA control组(P<0.01)。p-EGFP-C1、p-EGFP-C1/Smad1、siRNA control、Smad1 siRNA组细胞中Akt蛋白表达水平没有变化。p-EGFP-C1/Smad1组细胞MMP-9、MMP-2、p-Akt蛋白表达水平明显低于p-EGFP-C1组(P<0.01)。Smad1 siRNA组细胞MMP-9、MMP-2、p-Akt蛋白表达水平明显高于siRNA control(P<0.01)。胃腺癌细胞与Akt信号通路抑制剂作用后细胞增殖和迁移趋势与p-EGFP-C1/Smad1组一致。结论:Smad1在胃腺癌组织中低表达。Smad1能够抑制胃腺癌细胞增殖和迁移,作用机制与Akt信号通路有关。

Objective ：To investigate the expression of Smadl in gastric carcinoma and the influence on the migration ability of gastric cancer cells. Methods：Collected the protein from the gastric cancer tissues and corresponding adjacent tissues, the expression level of Smadl was detected by Western blot. In HGC-27 gastric cancer cells as the research object, the carrier cells transfected with overexpression of Smadl （p-EGFP-C1/Smadl） and Smadl small interfering RNA （Smadl siRNA）,while transfection of p-EGFP-C1 and siRNA control as control. MTT to detect cell proliferation. Cell migration ability was detected with cell scratch test. The expression levels of MMP-9, MMP-2, p-Akt and Akt in cells were detected by Western blot. Akt signal pathway inhibitor LY294002 （20 μg/ml） in gastric adenocarcinoma cells, MTT for cell proliferation, cell scratch assay for cell migration. The expression levels of MMP-9, MMP-2, p- Akt,Akt were detected by Western blot. Results： Smadl in gastric carcinoma was significantly lower than the adjacent tissues （P〈 0. 01 ）. The cell survival rate and migration rate of p-EGFP-C1/Smadl group were significantly lower than that of p-EGFP-C1 group （P〈0. 01 ）. The cell survival rate and migration rate of Smadl siRNA group were significantly higher than those in the siRNA control group （P〈0. 01 ）. The expression levels of Akt protein in P-EGFP-C1 ,p-EGFP-C1/Smadl, Smadl siRNA, siRNA control cells did not change. The expression levels of MMP-9 ,MMP-2 and p-Akt in p-EGFP-C1/Smadl group were significantly lower than that in p-EGFP- C1 group （P〈0. 01 ）. The expression levels of MMP-9, MMP-2 and p-Akt in Smadl siRNA group were significantly higher than that in control siRNA （P〈0.01）. The cell proliferation and migration trends in gastric cancer cells effected by Akt signaling pathway inhibitor consistent with the p-EGFP-C1/Smadl group. Conclusion：Low expression of Smadl in gastric cancer tissue. Smadl can inhibit the pro- liferation and migration of gastric cancer cells, the mechanism of action is related to the Akt signaling pathway.