miR-299-3p靶向ALDH3A1下调NF-κB信号通路诱导胃癌细胞凋亡的机制研究

The effects of miR-299-3p on the apoptosis of gastric cancer SGC-7901 cells by targetedly down-regulating NF-κB signaling pathway

目的研究miR-299-3p对胃癌SGC-7901细胞凋亡的影响及潜在的分子机制。方法以胃正常上皮细胞GES-1为对照,qRT-PCR检测胃癌细胞株SGC-7901、MKN-45和BGC-823中ALDH3A1 mRNA和miR-299-3p的表达,Western blot检测ALDH3A1、凋亡蛋白Bax和Bcl2以及NF-κB信号通路蛋白IκB-α、p-IκB-α和NF-κB的表达水平,流式细胞术检测细胞凋亡,双荧光素酶报告系统验证miR-299-3p和ALDH3A1的关系。结果与对照相比,胃癌细胞株SGC-7901、MKN-45和BGC-823中miR-299-3p表达量降低(P<0.05),ALDH3A1的mRNA和蛋白表达量升高(P<0.05);过表达miR-299-3p可促进Bax表达,抑制Bcl2表达,抑制NF-κB信号通路,从而诱导SGC-7901细胞凋亡;抑制ALDH3A1表达可诱导SGC-7901细胞凋亡,miR-299-3p靶向负调控ALDH3A1的表达;过表达ALDH3A1逆转了上调miR-299-3p对SGC-7901细胞凋亡及NF-κB信号通路的影响。结论miR-299-3p通过靶向ALDH3A1下调NF-κB信号通路诱导胃癌SGC-7901细胞凋亡。miR-299-3p是胃癌的潜在分子靶点。

Objective To investigate the effects of miR-299-3p on the apoptosis of gastric cancer SGC-7901 cells,and to explore its possible molecule mechanism.Methods The normal gastric epithelial cells GES-1 were taken as control group.The expression levels of ALDH3A1 mRNA and miR-299-3p in gastric cancer cell lines SGC-7901,MKN-45 and BGC-823 were detected by qRT-PCR,and the expression levels of ALDH3A1,apoptosis-related proteins Bax and Bcl2,and NF-κB signaling pathway proteins-IκB-α,p-IκB-αand NF-κB were detected by Western Blot,and the apoptosis of SGC-7901 cells was detected by flow cytometry,and the correlation between miR-299-3p and ALDH3A1 was verified by using dual-luciferase reporter assay.Results Compared with those in control group,the expression levels of miR-299-3p in gastric cancer cell lines SGC-7901,MKN-45 and BGC-823 were decreased significantly(P<0.05),however,the expression levels of ALDH3A1 mRNA and protein were significantly increased(P<0.05).The overexpression of miR-299-3p promoted the expression of Bax,inhibited the expression of Bcl2,and inhibited the signal pathway of NF-κB,thereby inducing the apoptosis of SGC-7901 cells.To inhibit ALDH3A1 could induce the apoptosis of SGC-7901 cells.The expression of ALDH3A1 was negatively regulated by miR-299-3p.In addition the overexpression of ALDH3A1 reversed the effects of up-regulation of miR-299-3p on the apoptosis and NF-κB signaling pathway of SGC-7901 cells.Conclusion The miR-299-3p can induce the apoptosis of gastric cancer SGC-7901 cells by targeting ALDH3A1 to down-regulate NF-κB signaling pathway,and miR-299-3p is a potential molecular target of gastric cancer.