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目的探讨全反式维甲酸(ATRA)对人胃腺癌SGC7901细胞的增殖、凋亡和迁移影响及其可能机制,同时分析ATRA对胞膜蛋白caveolin-1表达和定位的影响。方法 MTT法检测ATRA在不同浓度(5、10、25、50、100μmol/L)和不同时间(24、48、72 h)对人胃腺癌SGC7901细胞增殖的影响;Ho-chest33258染色实验观察ATRA对人胃腺癌SGC7901细胞凋亡的影响;细胞划痕实验观察AT-RA对人胃腺癌SGC7901细胞迁移的影响;Westernblot分析ATRA对人胃腺癌SGC7901细胞凋亡相关蛋白、ERK磷酸化水平和caveolin-1表达的影响;免疫荧光法检测caveolin-1在人胃腺癌SGC7901细胞中的表达和定位。结果 MTT实验显示ATRA可明显抑制人胃腺癌SGC7901细胞增殖。Hochest33258染色实验显示ATRA促进人胃腺癌SGC7901细胞凋亡。细胞划痕实验示ATRA可明显抑制人胃腺癌SGC7901细胞迁移。Western blot实验表明:①ATRA降低人胃腺癌SGC7901细胞的凋亡抑制蛋白Bcl-2的表达,而对凋亡促进蛋白Bax和酶原活性的caspase-3无明显影响;②ATRA抑制人胃腺癌SGC7901细胞的ERK磷酸化水平:从45 min开始下降,到2 h达到最大抑制,随后抑制作用逐渐减弱;③ATRA处理人胃腺癌SGC7901细胞后caveo-lin-1表达没有影响,但胞浆内caveolin-1含量明显降低,胞膜上caveolin-1表达明显增加。免疫荧光实验发现ATRA促进人胃腺癌SGC7901细胞的caveo-lin-1在细胞膜上定位。结论 ATRA可明显抑制人胃腺癌SGC7901细胞增殖,促进细胞凋亡,抑制细胞迁移,降低ERK的磷酸化水平,增加caveolin-1在细胞膜上定位。
Objective To investigate the effect of all-trans retinoic acid (ATRA) on the proliferation, apoptosis and migration of human gastric adenocarcinoma SGC7901 cells and its possible mechanism, and to analyze the effect of ATRA on the expression and localization of caveolin-1. Methods MTT assay was used to detect the effect of ATRA on the proliferation of human gastric adenocarcinoma SGC7901 cells at different concentrations (5, 10, 25, 50, 100 and 100 micromol / L) The effect of AT-RA on the migration of human gastric adenocarcinoma SGC7901 cells was observed by cell scratch assay. The effect of ATRA on apoptosis-related protein, ERK phosphorylation and caveolin-1 in human gastric adenocarcinoma SGC7901 cells was analyzed by Western blot analysis The expression and localization of caveolin-1 in human gastric adenocarcinoma SGC7901 cells were detected by immunofluorescence. Results MTT assay showed that ATRA could significantly inhibit the proliferation of human gastric adenocarcinoma SGC7901 cells. Hochest33258 staining experiments showed that ATRA promotes the apoptosis of human gastric adenocarcinoma SGC7901 cells. Cell scratch assay showed that ATRA could significantly inhibit the migration of human gastric adenocarcinoma SGC7901 cells. Western blot showed that: ①ATRA decreased the expression of Bcl-2, but had no effect on the activity of pro-apoptotic protein Bax and caspase-3; ②ATRA inhibited the proliferation of human gastric adenocarcinoma SGC7901 cells The level of ERK phosphorylation decreased from 45 min to the maximum at 2 h, and then the inhibition decreased gradually. ③ The expression of caveolin-1 in the cytoplasm of ATG-treated human gastric adenocarcinoma SGC7901 cells had no effect Decreased, the caveolin-1 expression on the membrane increased significantly. Immunofluorescence showed that ATRA could promote the localization of caveo-lin-1 in human gastric adenocarcinoma SGC7901 cells. Conclusion ATRA can significantly inhibit the proliferation of human gastric adenocarcinoma SGC7901 cells, promote cell apoptosis, inhibit cell migration, reduce the phosphorylation of ERK and increase the localization of caveolin-1 on the cell membrane.