STK17A基因经TGF-β/SMAD信号通路调控宫颈癌细胞增殖凋亡及侵袭的机制研究  

STK17A regulates proliferation,invasion and apoptosis of cervical cancer through targeting TGF-β/SMAD signaling pathway

作　　者：刘海霞[1] 宋梅[1,2] 戴淑凤[1] LIU Hai-Xia;SONG Mei;DAI Shu-Feng(The Third People′s Hospital of Qingdao,Qingdao 266041,China)

机构地区：[1]青岛市第三人民医院,青岛266041 [2]青岛市肿瘤医院,青岛266042

出　　处：《中国免疫学杂志》2020年第6期707-713,共7页Chinese Journal of Immunology

摘　　要：目的:探究丝氨酸/苏氨酸激酶17A(STK17A)基因介导转化生长因子β(TGF-β)/SMAD信号通路对宫颈癌细胞增殖、侵袭及凋亡的调控机制。方法:收集我院43例宫颈癌患者癌组织及癌旁组织。HE染色观察组织病理结构。免疫组化检测组织中STK17A阳性表达。筛选STK17A低表达宫颈癌细胞系并将细胞系分为阴性对照组(宫颈癌细胞转染含有无关序列的重组质粒)、基因沉默组(宫颈癌细胞转染含有STK17A shRNA的重组质粒)、基因过表达组(宫颈癌细胞转染含有STK17A片段的重组质粒)、通路抑制剂组(TGF-β/SMAD通路特异性抑制剂处理宫颈癌细胞)、联合组(TGF-β/SMAD通路抑制联合STK17A过表达处理细胞)。qRT-PCR和Western blot检测各组细胞STK17A、TGF-β1、SMAD3、E-cadherin、N-cadherin和TIMP3的表达。CCK-8检测各组细胞活力。细胞划痕和Transwell实验检测各组细胞迁移和侵袭能力。Annexin V-FITC/PI双染检测各组细胞凋亡情况。结果:与癌旁组织相比,宫颈癌组织排列紊乱,无极性(层次和结构)且STK17A低表达。细胞实验中:与阴性对照相比,过表达STK17A或抑制TGF-β/SMAD通路后,TGF-β1、SMAD3、E-cadherin mRNA和蛋白表达下调,但N-cadherin、TIMP3 mRNA和蛋白表达上调(均P<0.05)。同时,相对于阴性对照组,过表达STK17A或抑制TGF-β/SMAD通路后,24 h和48 h细胞活力受到明显抑制、细胞侵袭转移能力显著降低且细胞凋亡被显著诱导(均P<0.05)。结论:STK17A过表达可能通过抑制TGF-β/SMAD信号通路的活化进而抑制宫颈癌细胞增殖、侵袭同时诱导其凋亡,STK17A有望成为宫颈癌治疗的潜在重要靶点。Objective:To elucidate the underlying mechanism by which(Serine/threonine kinase 17A,STK17A)affect proliferation,apoptosis and invasion of cervical cancer cells through TGF-β/SMAD signaling pathway.Methods:Cancer tissues and paracancerous tissues from 43 cases of cervical cancer patients were collected.HE staining was utilized to detect the histopathological structure.Positive expression rate of STK17A was measured by using immunohistochemistry.The cell line with lowest expression of STK17A was selected and divided into negative control group(cervical cancer cells transfected with plasmids containing irrelevant sequences);STK17A shRNA group(cervical cancer cells transfected with plasmids containing STK17A shRNA);STK17A mimic group(cervical cancer cells transfected with plasmids containing STK17A mimic);TGF-β/SMAD inhibitor group(cervical cancer cells treated with TGF-β/SMAD inhibitor);STK17A mimic+TGF-β/SMAD inhibitor group(cervical cancer cells transfected with plasmids containing STK17A mimic and treated with TGF-β/SMAD inhibitor).qRT-PCR and Western blot were used to determine the mRNA and protein expression of STK17A,TGF-β1,SMAD3,E-cadherin,N-cadherin and TIMP3 in each group of cells.CCK8 was utilized to measure the cell viability in each group.Cell scratch test and Transwell assay were adopted to detect the migration and invasion ability in each group.Annexin V-FITC/PI staining was used to determine cell apoptosis in each group.Results:Compared with paracancerous tissue,arrangement of cervical cancer tissue was disordered,the tissue had non-polar(hierarchy and structure).At the same time,STK17A expression in cervical cancer tissue was inhibited compared with paracancerous tissue.As for the cell experiments,compared with the negative control(NC)group,mRNA and protein expression of TGF-β1,SMAD3,E-cadherin were decreased while N-cadherin,TIMP3 mRNA and protein expression were increased when cervical cancer cells were transfected with plasmids containing STK17A mimic or treated with TGF-β/SMAD inhibitor(

关 键 词：丝氨酸/苏氨酸激酶17A 宫颈癌 TGF-Β/SMAD信号通路 增殖 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]