SIRT1沉默对人甲状腺乳头状癌细胞株TPC-1生物学功能的影响  

Effect of Silencing SIRT1 on the Biological Behavior of the Human Thyroid Papillary Carcinoma Cell Line TPC-1

作　　者：彭华童 吴文艺[1] 张丽婷[2] 林建清[1] 余艺煌 吴春林[1] 黄种心[1] 邱建龙[3] PENG Huatong;WU Wenyi;ZHANG Liting;LIN Jianqing;YU Yihuang;WU Chunlin;HUANG Zhongxin;QIU Jianlong(Department of Oncological Surgery,The Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Endocrinology,The 180th Military Hospital of Chinese People’s Liberation Army,Quanzhou 362000,China;Department of Pathology,The 180th Military Hospital of Chinese People’s Liberation Army,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院肿瘤外科,福建泉州362000 [2]解放军第一八零医院内分泌科,福建泉州362000 [3]解放军第一八零医院病理科,福建泉州362000

出　　处：《中国医科大学学报》2018年第4期321-328,共8页Journal of China Medical University

基　　金：福建省卫生系统中青年骨干人才培养项目(2015-ZQNZD-26)

摘　　要：目的 观察沉默信息调节因子2相关酶1(SIRT1)基因沉默对人甲状腺乳头状癌细胞株TPC-1生物学功能的影响。方法 利用SIRT1siRNA技术下调SIRT1在甲状腺乳头状癌细胞株TPC-1中的表达,应用q RT-PCR方法检测siRNA的干扰效率,筛选出干扰效率最强的siRNA,并用Western blotting验证SIRT1蛋白的表达,利用CCK8法、β-半乳糖苷酶染色法、Hoechst染色法及流式细胞技术检测转染细胞的增殖抑制率、衰老、凋亡与周期的影响。结果 转染SIRT1siRNA的甲状腺乳头状癌细胞较未转染细胞SIRT1 m RNA及蛋白表达水平显著降低(P<0.05);转染细胞增殖率显著低于未转染细胞(P<0.05),转染细胞增殖明显受到抑制(P<0.05);转染组衰老细胞比例明显增加(P<0.05),凋亡细胞比例明显升高(P<0.05);转染细胞出现G1期停滞(67.99%±0.87%,P<0.05)。结论 SIRT1在人甲状腺乳头状癌细胞株TPC-1的低表达能抑制细胞的增殖能力,阻滞细胞周期,并诱导细胞的衰老及凋亡。Objective To investigate the effect of silencing SIRT1 on the biological behavior of the human thyroid papillary carcinoma cell line TPC-1.Methods SIRT1 siRNA was used to downregulate the expression of SIRT1 in TPC-1.The interference efficiency of siRNA was detected using qRT-PCR.The siRNA with the strongest interference efficiency was selected and the expression of SIRT1 protein was verified by Western blotting.The inhibitory rate of proliferation,senescence,apoptosis,and cycling of transfected cells was detected using the CCK8 method,β-galactosidase staining,Hoechst staining,and flow cytometry,respectively.Results The SIRT1 expression in thyroid papillary carcinoma cells transfected with siRNA was lower(P<0.05)than that in the untransfected cells.The proliferation of transfected cells was significantly inhibited(P<0.05)and their proliferation rate was lower than that of untransfected cells(P<0.05).The proportion of aged cells and the percentage of apoptotic cells in the transfected carcinoma cells was significantly increased(P<0.05).The transfection of SIRT1 siRNA increased the number of cells in the G1 phase(67.99%±0.87%,P<0.05).Conclusion Reduced SIRT1 expression in the human thyroid papillary carcinoma cell line TPC-1 can inhibit cell proliferation,block the cell cycle,and induce cell senescence and apoptosis.

关 键 词：沉默信息调节因子2相关酶1 RNA干扰 甲状腺乳头状癌 

分 类 号：R653[医药卫生—外科学]