RNA干扰FLOT2基因表达下调NF-κB信号对胃癌细胞凋亡诱导作用研究

FLOT2 gene expression with RNA interference induces apoptosis in gastric cancer

目的 探讨RNA干扰FLOT2基因表达对胃癌细胞凋亡及NF-κB信号的影响。方法 Western blotting检测人胃癌BGC823、SGC7901和MKN28细胞中FLOT2蛋白表达。BGC823细胞分为空白组、阴性组和si-FLOT2组,参照脂质体Lipofectamine TM 2000将siRNA转染细胞,细胞转染48 h,Western blotting检测FLOT2、NF-κB p65、IKK-β、p-IKK-β和Bax的蛋白表达。流式细胞术检测细胞凋亡率。结果 BGC823、SGC7901和MKN28胃癌细胞中FLOT2蛋白表达均显著高于人正常胃黏膜上皮细胞GES1(P<0.05 )。转染si-FLOT2的BGC823细胞FLOT2蛋白表达显著低于空白组(P<0.05)。与空白组比较,si-FLOT2组细胞凋亡率显著升高,NF-κB p65、IKK-β和p-IKK-β蛋白表达显著降低,Bax蛋白表达显著升高(P<0.05)。结论 RNA干扰FLOT2基因表达可诱导胃癌细胞凋亡,机制可能与下调NF-κB信号通路有关。

Objective To investigate the effect of FLOT2 gene expression with RNA interference on apoptosis and NF-κB signaling in gastric cancer cells.Methods Western blotting was used to detect the expression of FLOT2 protein in human gastric cancer BGC823, SGC7901 and MKN28 cells. BGC823 cells were divided into blank group, negative group and si-FLOT2 group. siRNA was transfected into cells according to the liposome LipofectamineTM2000. The cells were transfected for 48 hours, expressions of FLOT2, NF-κB p65, IKK-β, p-IKK-β and Bax proteins were detected by Western blotting. The cell apoptosis rate was detected by flow cytometry.Results The expression of FLOT2 protein in BGC823, SGC7901 and MKN28 gastric cancer cells was significantly higher than that in normal gastric mucosal epithelial cells(P<0.05). The expression of FLOT2 protein in BGC823 cells transfected with si-FLOT2 was significantly lower than that in blank group(P<0.05). Compared with the blank group, the apoptosis rate in si-FLOT2 group was significantly increased, and the expressions of NF-κB p65, IKK-β and p-IKK-β proteins were significantly decreased, and the expression of Bax protein was significantly increased(P<0.05).Conclusion FLOT2 gene expression with RNA interference can induce apoptosis in gastric cancer cells. The mechanism may be related to down regulation of NF-κB signaling pathway.