SGK1调控NF-κB通路影响宫颈癌细胞增殖和凋亡的机制研究

The expression of SGK1 protein in cervical cancer and its mechanism of regulating NF-κB pathway affecting the proliferation and apoptosis of cervical cancer cells

目的探讨血清和糖皮质激素调节蛋白激酶1(SGK1)在宫颈癌患者组织中的表达情况,以及对宫颈癌细胞Caski增殖和凋亡的影响和可能的分子机制。方法采用蛋白免疫印迹法(Western blot)检测SGK1蛋白在人正常宫颈组织和宫颈癌组织中的表达情况,以20μmol/L的SGK1抑制剂GSK650394处理人宫颈癌Caski细胞,实验分为Control组(对照)、GSK组(20μmol/L SGK1抑制剂GSK650394)和GSK+PMA组(20μmol/L SGK1抑制剂GSK650394和1μmol/L NF-κB信号通路激活剂PMA),噻唑蓝比色法(MTT)和流式细胞术分析Caski细胞体外增殖和凋亡情况,Western blot分析各组Caski细胞中SGK1、NF-κB和NF-κB抑制蛋白α(IκBα)的表达水平。结果SGK1在宫颈癌组织中的表达显著高于正常宫颈组织(P<0.05),与Control组比较,GSK组Caski细胞中SGK1蛋白表达明显受到抑制,增殖活力明显降低,凋亡率明显升高,NF-κB和IκBα的表达水平明显降低(P均<0.05);与GSK组比,GSK+PMA组Caski细胞中SGK1蛋白表达上调,增殖活力升高,凋亡率降低,NF-κB和IκBα的表达水平升高(P均<0.05)。结论SGK1蛋白在宫颈癌组织中的表达异常升高,抑制SGK1蛋白能够阻碍宫颈癌Caski增殖并诱导细胞凋亡,其作用机制可能与阻断NF-κB信号通路的激活有关。

Objective To investigate the expression of serum-and glucocorticoid-regulated protein kinase 1(SGK1)in the tissues of cervical cancer patients,as well as the influence on the proliferation and apoptosis of cervical cancer cells Caski and the possible molecular mechanism.Methods Western Blot was used to detect the expression of SGK1 protein in human normal cervical tissues and cervical cancer tissues,and 20μmol/L SGK1 inhibitor GSK650394 was used to treat human cervical cancer Caski cells.They were divided into Control group(control),GSK group(20μmol/L SGK1 inhibitor GSK650394)and GSK+PMA group(20μmol/L SGK1 inhibitor GSK650394 and 1μmol/L NF-κB signaling pathway activator PMA).Thiazole blue colorimetry(MTT)and flow cytometry were used to analyze the proliferation and apoptosis of Caski cells in vitro.Western blot was used to analyze the expression levels of SGK1,NF-κB and NF-κB inhibitory protein alpha(IκBα)in Caski cells of each group.Results The expression levels of SGK1 in cervical cancer tissues were significantly higher than those in normal cervical tissues(P<0.05).Compared with those in control group,the SGK1 protein expression levels in Caski cells of the GSK group were significantly inhibited,the proliferation activity was significantly reduced,the apoptosis rate was significantly increased,and the expression levels of NF-κB and IκBαwere significantly reduced(P<0.05).Compared with those in GSK group,the expression levels of SGK1 protein in Caski cells in GSK+PMA group were up-regulated,and proliferation activity was increased,but,apoptosis rate was decreased,and the expression levels of NF-κB and IκBαwere increased(P<0.05).Conclusion The expression levels of SGK1 protein in cervical cancer tissues are abnormally increased.To inhibit the expression of SGK1 protein can suppress the proliferation of cervical cancer Caski and induce cell apoptosis,and its action mechanism may be related to blocking the activation of NF-κB signaling pathway.