干扰NDRG1表达通过调控PI3K/Akt信号通路对皮肤鳞状细胞癌细胞凋亡的影响  

Silencing NDRG1 Expression Affects the Apoptosis of Skin Squamous Cell Carcinoma Cells by Regulating the PI3K/Akt Signaling Pathway

作　　者：林简 刘晓红 王荣坤 李涛 李娜 陈惠娟 LIN Jian;LIU Xiaohong;WANG Rongkun;LI Tao;LI Na;CHEN Huijuan(Dermatology&STD Department,Sanya People's Hospital,Sanya 572000 China)

机构地区：[1]三亚市人民医院皮肤性病科,海南三亚572000

出　　处：《中国皮肤性病学杂志》2020年第9期1005-1011,共7页The Chinese Journal of Dermatovenereology

摘　　要：目的探讨小干扰RNA(siRNA)干扰N-myc下游调节基因1(NDRG1)基因表达对皮肤鳞状细胞癌A431细胞凋亡及PI3K/AKT信号通路的影响。方法将siRNA-NDRG1干扰质粒转染至人皮肤鳞状细胞癌A431细胞中,采用RT-PCR和Western blot检测A431细胞中NDRG1 mRNA和蛋白表达;采用CCK-8、流式细胞术分别检测A431细胞增殖活性和凋亡率;采用Western blot检测A431细胞中凋亡相关蛋白Caspase3、Bax和Bcl-2以及PI3K/AKT信号通路相关蛋白PI3K、p-PI3K、AKT和p-AKT表达水平。采用100μg/L的PI3K/AKT信号通路特异性激活剂IGF-1与si-NDRG1单独或联合处理A431细胞,流式细胞术和Western blot检测细胞凋亡率以及PI3K、p-PI3K、AKT和p-AKT蛋白表达水平。结果NDRG1 siRNA干扰可显著抑制A431细胞中NDRG1 mRNA和蛋白表达;干扰NDRG1基因表达可显著抑制A431细胞增殖活性,提高细胞凋亡率,并上调Cleaved-Caspase3和Bax蛋白表达水平,下调Bcl-2、p-PI3K及p-AKT蛋白水平;然而,IGF-1与si-NDRG1联合作用时,IGF-1可抑制NDRG1基因干扰后对A431细胞凋亡的促进作用以及对p-PI3K和p-AKT蛋白的下调作用。结论干扰NDRG1基因表达可通过抑制PI3K/AKT信号通路活化诱导人皮肤鳞状细胞癌细胞凋亡。Objective To investigate the effects of small interfering RNA(siRNA) on NDRG1 gene expression on apoptosis and PI3 K/AKT signaling in A431 cutaneous squamous cell carcinoma.Methods siRNA-NDRG1 interfering plasmid was transfected into A431 cells.The expression levels of NDRG1 mRNA and protein in A431 cells were detected by RT-PCR and Western blot respectively.Proliferation activity and apoptosis rate of A431 cells were detected by CCK-8 and flow cytometry respectively.Western blot was used to detect the expression levels of apoptosis-related proteins Caspase3,Bax,Bcl-2 and PI3 K/AKT signaling proteins PI3 K,p-PI3 K,AKT and p-AKT in A431 cells.Treating with 100 μg/L PI3 K/AKT signaling pathway specific activator IGF-1 and si-NDRG1 alone or in combination in A431,the apoptosis rate was dected by flow cytometry,and the expression levels of PI3 K,p-PI3 K,AKT and p-AKT was determined by Western blot.Results NDRG1 siRNA interference significantly inhibits the expression of NDRG1 mRNA and protein in A431 cells.Interference with NDRG1 gene expression significantly inhibits proliferation activity of A431 cells,improves apoptosis rate,up-regulates Cleaved-Caspase3 and Bax protein expression levels,and down-regulates Bcl-2,p-PI3 K and p-AKT protein levels.However,when IGF-1 is combined with si-NDRG1,IGF-1 inhibits the apoptosis rate of A431 cells and increases protein levels of p-PI3 K and p-AKT after interference of NDRG1 gene.Conclusion Interference with NDRG1 gene expression induces apoptosis of human skin squamous cancer cells by inhibiting the PI3 K/AKT signaling pathway.

关 键 词：皮肤鳞状细胞癌 NDRG1基因 小干扰RNA PI3K/AKT信号通路 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤]