摘要
目的观察转染Survivin shRNA的皮肤鳞状细胞癌(c SCC)细胞系凋亡、增殖及NF-κB信号通路相关蛋白的表达变化。方法取Survivin mRNA相对表达量最高的c SCC细胞系A431细胞分为sh-Survivin组和sh-NC组,sh-Survivin组细胞转染Survivin shRNA,sh-NC组细胞转染NC shRNA。两组细胞转染48 h后,采用细胞周期实验检测各组细胞凋亡率。两组细胞转染第1、2、3、4 d时,采用MTS法检测两组细胞增殖能力(以OD值表示)。两组细胞转染48 h后,采用Western Blotting法检测两组细胞NF-κB抑制蛋白α(IκBα)、磷酸化IκB激酶α/β(pIKKα/β)、磷酸化p65(p-p65)蛋白。结果sh-Survivin组、sh-NC组细胞凋亡率分别为19.21%±2.54%、4.52%±0.31%,两组相比,P<0.05。sh-Survivin组细胞第1、2、3、4 d时的OD值分别为0.27±0.01、0.32±0.04、0.37±0.03、0.54±0.08,sh-NC组分别为0.27±0.02、0.38±0.03、0.67±0.05、0.88±0.07,其中sh-Survivin组细胞第3、4 d时的OD值显著低于sh-NC组(P均<0.05)。sh-Survivin组中IκBα、p-IKKα/β、p-p65蛋白的相对表达量分别为0.61±0.11、0.13±0.06、0.09±0.05,sh-NC组分别为0.11±0.04、0.76±0.15、0.81±0.19,两组相比,P均<0.05。结论转染Survivin shRNA的c SCC细胞系凋亡率升高、增殖能力下降、NF-κB信号通路受到抑制;干扰Survivin可能通过抑制NF-κB信号通路促进c SCC细胞凋亡、抑制c SCC细胞增殖。
Objective To observe the apoptosis,proliferation and expression of NF-κB signaling pathway-related proteins in the skin squamous cell carcinoma(c SCC)cell lines transfected with Survivin shRNA.Methods A431 cells with the highest relative expression of Survivin mRNA were divided into the sh-Survivin group and sh-NC group.Survivin shRNA was transfected into cells of the sh-Survivin group,and NC shRNA was transfected into cells of the sh-NC group.Cell cycle assay was used to detect apoptosis rate of each group at 48 h after transfection.At the 1 st,2 nd,3 rd and 4 th days of transfection,the proliferation ability of each group was detected by MTS method(represented by OD value).After transfection for 48 h,NF-κB inhibitory protein(IκBα),phosphorylated IκB kinaseα/β(p-IKKα/β)and phosphorylated p65(pp65)were detected by Western blotting.Results The apoptosis rates of the sh-Survivin group and sh-NC group were19.21%±2.54%and 4.52%±0.31%,respectively,with statistically significant difference(P<0.05).OD values of the sh-Survivin group were 0.27±0.01,0.32±0.04,0.37±0.03,and 0.54±0.08 on day 1,2,3,and 4,respectively,and those of sh-NC group were 0.27±0.02,0.38±0.03,0.67±0.05,and 0.88±0.07,respectively.OD value of the sh-Survivin group on day 3 and 4 were significantly lower than those of the sh-NC group(all P<0.05).The relative expression of IκBα,p-IKKα/βand p-p65 in the sh-Survivin group were 0.61±0.11,0.13±0.06 and 0.09±0.05,respectively,and those in the sh-NC group were 0.11±0.04,0.76±0.15 and 0.81±0.19,respectively,with statistically significant difference between every two groups(all P<0.05).Conclusion The apoptosis rate of c SCC cells transfected with Survivin shRNA increases,proliferation ability decreases,and NF-κB signaling pathway is inhibited;interfering Survivin may promote apoptosis and inhibit proliferation of c SCC cells by inhibiting NF-κB signaling pathway.
作者
谭亚琦
徐文俊
何焱玲
TAN Yaqi;XU Wenjun;HE Yanling(Beijing Chaoyang Hospital Affiliated to Capital Medical University,Beijing 100020,China)
出处
《山东医药》
CAS
2020年第21期15-18,共4页
Shandong Medical Journal
基金
北京市首都卫生发展科研专项项目(2017-3-3212)。
