siRNA干扰CTHRC1可体外抑制甲状腺乳头状癌TPC-1细胞的增殖并诱导凋亡  

Effects of RNA interference of CTHRC1 on proliferation and apoptosis of thyroid papillary cancer TCP-1 cells in vitro

作　　者：唐振宁[1] 丁小云 秦少杰[1] 张朝林[1] TANG Zhenning;DING Xiaoyun;QIN Shaojie;ZHANG Chaolin(Department of Oncology Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院肿瘤外三科,宁夏银川750004

出　　处：《南方医科大学学报》2021年第4期549-554,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81360390);宁夏自然科学基金(NZ13161,2019AAC03198);宁夏医科大学科学研究基金资助重点项目(XZ2015029)。

摘　　要：目的探讨胶原三股螺旋重复蛋白1(CTHRC1)甲状腺乳头状癌TPC-1细胞增殖、凋亡的影响。方法成功筛选干扰甲状腺乳头状癌TPC-1细胞CTHRC1表达的siRNA,以脂质体转染法将siRNA转染甲状腺乳头状癌TPC-1细胞,采用real-time PCR和Western blot检测转染后TCP-1细胞中CTHRC1mRNA和蛋白表达的变化。实验分组:将筛选的干扰序列转染TPC-1细胞作为干扰组(si-CTHRC1组),将随机阴性对照序列转染的TPC-1细胞作为阴性对照组(si-NC组),以不做任何处理TPC-1细胞为空白组(WT组)。采用CCK-8实验检测细胞增殖;流式细胞术AV/PI双染观察各组TCP-1细胞凋亡的变化;采用Western blot法检测各组TPC-1细胞中含剪切型半胱氨酸的天冬氨酸蛋白水解酶-3(c-Caspase-3)蛋白、剪切型多聚ADP核糖聚合酶1(c-PARP1)蛋白表达水平以及磷酸化细胞外信号调节激酶1/2(ERK1/2)表达水平。结果将筛选出明显抑制TPC-1细胞CTHCR1表达的siRNA转染细胞后,TPC-1细胞中CTHRC1的mRNA和蛋白表达水平降低(P<0.05)。与WT组和si-NC组相比,si-CTHRC1组的细胞活降低(P<0.05);与对照组相比,si-CTHRC1组的TPC-1细胞细胞凋亡率升高,Western blot实验发现:si-CTHRC1组的c-caspase-3、c-PARP1蛋白表达水平均升高(P<0.05);si-CTHRC1组ERK1/2蛋白的磷酸化水平增加。结论干扰CTHRC1能够抑制TPC-1细胞的增殖并诱导凋亡,此过程可能通过激活ERK1/2信号通路介导。Objective To explore the role of CTHRC1 in regulating the proliferation and apoptosis of papillary thyroid cancer cells.Methods Papillary thyroid cancer TPC-1 cells were transfected with a small interfering RNA(siRNA)targeting CTHRC1,with the cells transfected with a scrambled sequence as the negative control.The changes in cell proliferation and apoptosis were assessed using cell counting kit-8(CCK-8)and flow cytometry with AV/PI double staining,respectively.The expression of c-caspase-3,c-PARP1 and phosphorylation of ERK1/2 in the cells were examined with Western blotting.Results Transfection with the siRNA sequence significantly decreased the mRNA and protein levels of CTHRC1 in TCP-1 cells(P<0.05).Compared with blank and negative control cells,TCP-1 cells with RNA interference of CTHRC1 showed significantly lowered proliferative activity and enhanced cell apoptosis(P<0.05)with significantly increased expressions of c-caspase-3 and c-PARP1 and phosphorylation of ERK1/2(P<0.05).Conclusion RNA interference of CTHRC1 promotes the proliferation and inhibits apoptosis of papillary thyroid cancer cells possibly by activating the ERK1/2 pathway.

关 键 词：甲状腺癌 CTHRC1 增殖 凋亡 SIRNA干扰 

分 类 号：R736.1[医药卫生—肿瘤]