KDR调节PI3K/AKT/mTOR信号通路对宫颈癌增殖、迁移和侵袭的影响  

Influences of KDR on the proliferation, migration and invasion of cervical cancer by regulating PI3K/AKT/mTOR signaling pathway

作　　者：李颖 王凯 胡莉 LI Ying;WANG Kai;HU Li(Department of Obstetrics and Gynecology,Affiliated Hospital of Zunyi Medical University,Zunyi,Guizhou 563000,China)

机构地区：[1]遵义医科大学附属医院妇产科,贵州遵义563000

出　　处：《中国优生与遗传杂志》2022年第12期2113-2120,共8页Chinese Journal of Birth Health & Heredity

基　　金：遵义市联合基金项目:遵市科合HZ字(2019)88号。

摘　　要：目的 探讨激酶插入区受体(KDR)调节磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对宫颈癌增殖、迁移和侵袭的影响。方法 通过实时荧光定量PCR实验和免疫组织化学染色测定人宫颈癌组织和其癌旁组织中KDR的表达。体外培养HeLa细胞,随机分为对照组、KDR siRNA组、KDR siRNA阴性对照组、KDR siRNA+740 Y-P(PI3K激活剂)组,分组转染后,通过MTT实验、平板集落形成实验检测各组HeLa细胞增殖;通过Hoechst 33258染色检测各组HeLa细胞凋亡;通过划痕实验、Transwell实验分别检测各组HeLa细胞迁移及侵袭;通过免疫荧光染色检测各组HeLa细胞Bax、Bcl-2表达;通过免疫印迹实验检测各组HeLa细胞上皮间充质转化相关蛋白(Claudin-1、ZO-1、Vimentin、E-cadherin)、KDR蛋白及PI3K/AKT/mTOR通路相关蛋白(p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR)表达。结果 与癌旁组织相比,宫颈癌组织中KDR mRNA表达和KDR阳性表达率明显升高(P<0.05)。与对照组相比,KDRsiRNA组细胞增殖率、集落生成率、迁移距离、侵袭数、Claudin-1、Vimentin与KDR蛋白表达、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR降低(P<0.05),凋亡率、Bax/Bcl-2、ZO-1及E-cadherin蛋白表达升高(P<0.05);KDR siRNA阴性对照组细胞各指标无显著差异(P>0.05)。740 Y-P可逆转KDR siRNA对细胞各指标的作用。结论 敲低KDR可通过抑制PI3K/AKT/mTOR信号激活而促进宫颈癌细胞凋亡,降低其增殖、迁移和侵袭活性。Objective To investigate the influences of kinase insert domain receptor(KDR) on the proliferation, migration and invasion of cervical cancer by regulating phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR) signaling pathway. Methods The expression of KDR in human cervical cancer tissue and its adjacent tissue was determined by real-time quantitative PCR and immunohistochemical staining. HeLa cells were cultured in vitro and randomly grouped into control group, KDR siRNA group, KDR siRNA negative control group, and KDR siRNA+740Y-P(PI3K activator) group. After grouping and transfection, the proliferation of HeLa cells in each group was detected by MTT assay and plate colony formation assay. The apoptosis of HeLa cells in each group was detected by Hoechst 33258 staining. The migration and invasion of HeLa cells in each group were detected by scratch test and Transwell test respectively;the expressions of Bax and Bcl-2 in HeLa cells in each group were detected by immunofluorescence staining. The expressions of epithelial-mesenchymal transition-related proteins(Claudin-1, ZO-1, Vimentin, E-cadherin), KDR protein and PI3K/AKT/mTOR pathway-related proteins(p-PI3K/PI3K, p-AKT/AKT, p-mTOR/mTOR) were measured by western blotting.Results Compared with adjacent tissues, the expression of KDR mRNA and the positive expression rate of KDR in cervical cancer tissues was greatly increased(P<0.05). Compared with the control group, the cell proliferation rate, colony formation rate, migration distance, invasion number, Claudin-1, Vimentin and KDR protein expressions, p-PI3K/PI3K, p-AKT/AKT,p-mTOR/mTOR in KDR siRNA group decreased(P<0.05), the apoptosis rate, Bax/Bcl-2, ZO-1 and E-cadherin protein expressions increased(P<0.05). There was no obvious difference in each index of KDR siRNA negative control group(P>0.05).740 Y-P could reverse the effect of KDR siRNA on the above indexes. Conclusion KDR knockdown can inhibit the activation of PI3K/AKT/mTOR signaling to promote the apoptosis of

关 键 词：KDR PI3K/AKT/MTOR 宫颈癌 增殖 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]