地西他滨对胃癌细胞AGS增殖和侵袭的影响

Effect of decitabine on proliferation and invasion of gastric cancer AGS cells

目的探究地西他滨对胃癌AGS细胞增殖和侵袭的影响及其作用机制。方法本研究时间为2017年10月至2018年12月,采用地西他滨处理购自于中国科学院细胞库的胃癌AGS细胞,应用MTT法检测细胞增殖率,亚硫酸氢钠转化测序法检测APC基因启动子的甲基化程度,实时荧光定量PCR法检测APC基因mRNA的表达,Western blotting检测APC蛋白表达,细胞锚定非依赖性生长分析法和Transwell细胞侵袭法检测细胞的增殖和侵袭能力。结果AGS细胞中,APC基因启动子高度甲基化(85.7±5.4)%;地西他滨处理AGS细胞后,细胞活力受到抑制,且呈浓度和时间依赖性。APC基因启动子去甲基化,mRNA和蛋白表达水平均升高(P<0.05),5μM升高(2.3±0.1)倍,10μM升高(3.6±0.2)倍,25μM升高(4.1±0.1)倍,50μM升高(3.9±0.2)倍,增殖和侵袭能力均减弱(P<0.05),AGS组(290.3±5.7)个细胞,5μM为(213.2±6.7)个细胞,10μM为(160.4±7.3)个细胞,25μM为(145.8±6.2)个细胞,50μM为(119.2±8.6)个细胞。结论地西他滨可以抑制AGS细胞的增殖和侵袭,表现出抗肿瘤活性,其相关机制可能为去除AGS细胞中APC基因的甲基化,恢复APC基因的表达。

Objective To investigate the effect and mechanisms of decitabine on the proliferation and invasion of gastric cancer AGS cells.Methods The study time was from October 2017 to December 2018.The AGS cells which purchased from the Cell Bank ofthe Chinese Academy of Sciences were treated with decitabine and the cell proliferation rate was detected by MTT assay.The methylation status of the APC gene promoter was detected by bisulfite sequencing.The APC mRNA levels were measured by qRT-PCR,and theprotein levels were detected by western blot.The proliferation and invasion ability of the AGS cells were detected by anchorage-independent growth assay and Transwell invasion assay.Results The APC gene promoter was hypermethylated in AGS cells[(85.7±5.4)%].After decitabine treating the AGS cells,the cell viability was inhibited in a concentration and time-dependent manner,and the APC genepromoter was demethylated.The APC mRNA and protein expression levels were increased[5μM increased(2.3±0.1)times,10μM increased(3.6±0.2)times,25μM increased(4.1±0.1)times,50μM increased(3.9±0.2)times](P<0.05).And the proliferation and invasion ability of AGS cells were weakened[AGS group:(290.3±5.7)cells,5μM:(213.2±6.7)cells,10μM:(160.4±7.3)cells,25μM:(145.8±6.2)cells,50μM:(119.2±8.6)cells](P<0.05).Conclusion Decitabine can inhibit the proliferation and invasion of AGS cellsand exert its anticancer effects,the related mechanism may be to remove the methylation of the APC gene and restore its expression.