渥曼青霉素通过PI3K-AKT-mTOR信号通路抑制人皮肤鳞状细胞癌细胞增殖  

Targeting PI3K-AKT-mTOR by Wortmannin Inhibits Cell Growth in Human Skin Squamous Cell Carcinoma

作　　者：朱晓琴 陈红 ZHU Xiaoqin;CHEN Hong(Department of Integrated Chinese and Western Medical Oncology,Central Hospital of Enshi Autonomous Prefecture,Enshi 445000,China;Department of Critical Care Medicine,Central Hospital of Enshi Autonomous Prefecture,Enshi 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院中西医结合肿瘤科,湖北恩施445000 [2]恩施土家族苗族自治州中心医院重症医学科,湖北恩施445000

出　　处：《中国皮肤性病学杂志》2020年第1期21-25,共5页The Chinese Journal of Dermatovenereology

摘　　要：目的研究渥曼青霉素对人皮肤鳞状细胞癌A431细胞增殖、凋亡及PI3K-AKT-mTOR信号通路的影响。方法 CCK8法检测A431细胞增殖活性,Caspase-glot-3/-9活性测定试剂盒检测Caspase-3/9活性,Western blot实验分析Cleaved caspase-3、Cleaved PARP、p-P85(Y458)、p-AKT(S473和T308)和p-S6K(T389)蛋白表达水平,ELISA法检测单链DNA(ss DNA)的水平,流式细胞术检测A431细胞凋亡率。结果渥曼青霉素(0.5~6μmol/L)处理A431细胞48 h后,呈浓度和时间依赖性的抑制细胞增殖、诱导Caspase-3/9活性的增强、增高了细胞凋亡标志物ss DNA水平、还促使了促凋亡蛋白Cleaved caspase-3和Cleaved PARP的表达;4μmol/L的渥曼青霉素处理细胞48 h后,能明显诱导A431细胞的早期和晚期凋亡,还显著抑制了P85(Y458)、AKT(S473和T308)和S6K(T389)的磷酸化水平;渥曼青霉素相比于同浓度的PI3K-AKT-mTOR信号通路抑制剂(LY3023414、纳巴霉素、OSI-027和MK-2206)抗A431细胞增殖的活性更强。结论渥曼青霉素通过抑制PI3K-AKT-mTOR信号传导降低了人皮肤鳞状细胞癌A431细胞的增殖活性,诱导了细胞凋亡。Objective To study the effects of wortmannin on A431 cell proliferation,apoptosis and the activation of PI3 K-AKT-mTOR signaling pathway in human skin squamous cell carcinoma.Methods CCK8 assay was used to detect the proliferation activity of A431 cells,Caspase-3/9 activities were detected by Caspase-glot-3/-9 activity assay kits.Western blot was used to measure the expression levels of Cleaved caspase-3,Cleaved PARP,p-P85(Y458),p-AKT(S473 and T308)and p-S6 K(T389)proteins.ELISA assay was used to determine the level of single strand DNA(ss DNA).The apoptotic rate of A431 cells was also detected by flow cytometry.Results After treated with wortmannin(0.5~6μmol/L)for 48 h,it inhibited the proliferation of A431 cells,induced the activities of Caspase-3/9,increased the level of cell apoptotic marker ss DNA,and up-regulated the expression levels of Cleaved caspase-3 and Cleaved PARP in time-and dose-dependent manners.When A431 cells were treated with wortmannin(4μmol/L)for 48 h,the results showed that wortmannin could significantly induce the early and late apoptosis of A431 cells,and greatly inhibit the phosphorylation levels of P85(Y458),AKT(S473 and T308)and S6 K T389.Compared with other PI3 K-AKT-mTOR cascade inhibitors(LY3023414,rapamycin,OSI-027 and MK-2206),the anti-proliferation activity of wortmannin was much more stronger.Conclusion Wortmannin decreases the proliferation activity of human skin squamous cell carcinoma A431 cells and induces cell apoptosis via inhibiting the PI3 K-AKT-mTOR signaling pathway.

关 键 词：渥曼青霉素 A431细胞 人皮肤鳞状细胞癌细胞 PI3K-AKT-mTOR信号通路 

分 类 号：R739.5[医药卫生—肿瘤]