SOX1对甲状腺癌SW579细胞增殖和侵袭的影响及与Wnt通路的机制研究  

Effects of SOX1 on proliferation and invasion of thyroid cancer SW579 cells and mechanism of SOX1 with Wnt pathway

作　　者：喻建[1] 牟文辉[1] 向英豪 王波涌[2] YU Jian;MOU Wenhui;XIANG Yinghao(Department of General Surgery,People’s Hospital of Lichuan City,Hubei,Lichuan 445400,China)

机构地区：[1]湖北省利川市人民医院普外科,445400 [2]武汉大学中南医院甲乳外科

出　　处：《河北医药》2019年第17期2565-2570,共6页Hebei Medical Journal

摘　　要：目的探讨甲状腺癌SW579细胞中性别决定区Y框蛋白1(SOX1)表达及其抑制细胞增殖、侵袭的机制,并分析其与Wnt信号通路的作用关系。方法体外培养正常永生化表皮细胞Hacat、甲状腺癌SW579细胞,采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)及蛋白免疫印迹法(western Blot,WB)检测SOX1 mRNA及蛋白表达水平。实验分为空白对照组(未经任何处理的SW579细胞);阴性转染组(SOX1阴性对照质粒的SW579细胞);SOX1过表达组(含有SOX1过表达载体重组质粒的SW579细胞);Wnt激活剂组(Wnt通路激活剂的SW579细胞);联合组(SOX1重组质粒转染后加入Wnt通路激活剂的SW579细胞)。采用CCK-8法检测细胞增殖抑制率;Hoechst染色法检测细胞凋亡情况;Transwell法检测细胞迁移和侵袭;WB法检测β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(GSK-3β)及其磷酸化蛋白(p-GSK-3β)、细胞周期相关蛋白1(CyclinD1)、c-Myc蛋白表达情况。结果与正常组细胞比较,甲状腺癌SW579细胞中SOX1 mRNA及蛋白表达均显著降低(P<0.05);与空白对照组、阴性转染组、联合组比较,SOX1过表达组SW579细胞增殖抑制率、细胞凋亡率均显著升高(P<0.05),而Wnt激活剂组显著降低(P<0.05);与空白对照组、阴性转染组、联合组比较,SOX1过表达组SW579细胞迁移及侵袭数量、β-catenin、p-GSK-3β、CyclinD1、c-Myc蛋白表达水平均显著降低(P<0.05),而Wnt激活剂组显著升高(P<0.05)。结论SOX1可通过调控Wnt通路进而抑制甲状腺癌SW579细胞增殖及侵袭。Objective To investigate the expression of sex determining region Y-box protein 1(SOX1)in thyroid cancer SW579 cells and its mechanism of inhibiting cell proliferation and invasion,and to analyze its relationship with Wnt signaling pathway.Methods The normal immortalized epithelial cells Hacat and thyroid ccancer SW579 cells were cultured in vitro.The SOX1 mRNA and protein expression levels were detected by real-time quantitative PCR(qRT-PCR)and Western Blot(WB).The experiment was divided into blank control group(SW579 cells without any treatment),negative transfection group(SW579 cells with SOX1 negative control plasmid),SOX1 overexpression group(SW579 cells containing SOX1 overexpression vector recombinant plasmid),Wnt activator Group(SWV cells with Wnt pathway activator)and combination group(SW579 cells with Wnt pathway activator added after transfection of SOX1 recombinant plasmid).Inhibition rate of cell proliferation was detected by CCK8 method and apoptosis was detected by Hoechst staining.Cell migration and invasion were detected by Transwell method.Moreover the expression levels ofβcatenin,glycogen synthase kinase3β(GSK3β)and its phosphorylated protein(pGSK3β),cell cycle related protein 1(CyclinD1),cMyc protein were detected by WB method.Results As compared with those in normal group,the expression levels of SOX1 mRNA and protein in thyroid cancer SW579 cells were significantly decreased(P<0.05).As compared with those in blank control group,negative transfection group and combination group,the inhibition rate of cell proliferation and the apoptotic rate of SW579 cells were significantly increased in SOX1 overexpression group(P<0.05),which were significantly decreased in the Wnt activator group(P<0.05).Compared with the blank control group,the negative transfection group,and the combination group,the number of SW579 cell migration and invasion,and expression levels ofβcatenin,pGSK3β,CyclinD1,and cMyc protein were significantly decreased in the SOX1 overexpression group(P<0.05),while those indexes in

关 键 词：甲状腺癌 WNT通路 性别决定区Y框蛋白1 细胞增殖 细胞侵袭 

分 类 号：R736.1[医药卫生—肿瘤]