载脂蛋白C1高表达通过激活JAK2/STAT3信号通路促进甲状腺乳头状癌细胞的增殖并抑制凋亡  

High expression of apolipoprotein C1 promotes proliferation and inhibits apoptosis of papillary thyroid carcinoma cells by activating the JAK2/STAT3 signaling pathway

作　　者：宾禹 李子雯 左素微 孙思诺 李敏 宋佳茵 林旭 薛刚[2] 吴靖芳[1] BIN Yu;LI Ziwen;ZUO Suwei;SUN Sinuo;LI Min;SONG Jiayin;LIN Xu;XUE Gang;WU Jingfang(Laboratory of Morphology,Hebei North University,Zhangjiakou 075000,China;Department of Otolaryngology-Head and Neck Surgery,First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院形态学实验室,河北张家口075000 [2]河北北方学院附属第一医院耳鼻咽喉头颈外科,河北张家口075000

出　　处：《南方医科大学学报》2025年第2期359-370,共12页Journal of Southern Medical University

基　　金：河北省财政厅基金(ZF2023237);张家口市科技计划项目(2311004A)。

摘　　要：目的探讨载脂蛋白C1(APOC1)在甲状腺乳头状癌(PTC)组织中的表达及对PTC细胞增殖、凋亡及关键信号通路的影响。方法通过GEPIA 2及K-M数据库分析APOC1在PTC的表达及对预后的影响。采用免疫组织化学、Western blotting检测APOC1在PTC和癌旁组织中的表达以及3株PTC细胞和正常甲状腺Nthyori 3-1细胞的表达。采用细胞转染技术,构建APOC1敲低和过表达PTC细胞模型(TPC-1细胞和BCPAP细胞);采用Western blotting和RT-qPCR分别在蛋白水平和mRNA水平检验细胞模型构建是否成功。通过生长曲线、集落形成实验检测敲低和过表达APOC1后细胞的生长情况和集落形成能力;通过流式细胞术检测敲低和过表达APOC1对细胞周期、凋亡的影响。采用RT-qPCR和Western blotting检测增殖及凋亡相关靶基因P21,P27,CDK4,CyclinD1,BCL-2,Bax,caspase-3/caspase-9 mRNA和蛋白以及JAK2/STAT3信号通路关键蛋白的变化。结果成功构建APOC1敲低和过表达PTC细胞模型;PTC组织及3个PTC细胞株APOC1表达高于癌旁组织和Nthyori 3-1细胞(P<0.001)。与对照组相比,沉默组细胞增殖能力减弱(P<0.05);G0/G1期的细胞比例增高(P<0.01),S和G2期的细胞比例下降(P<0.05);细胞凋亡率明显增高(P<0.05);CDK4,CyclinD1,BCL-2 mRNA和蛋白表达量下调(P<0.05);p-JAK2,p-STAT3蛋白表达下调(P<0.001);增强组与沉默组结果相反:P21,P27,Bax,caspase-3/caspase-9 mRNA和蛋白表达下调(P<0.05);p-JAK2,p-STAT3蛋白表达上调(P<0.001)。JAK2抑制剂AG490显著减弱过表达APOC1对BCPAP细胞JAK2/STAT3信号通路的激活效应(P<0.01)。结论APOC1可能通过激活JAK2/STAT3信号通路,缩短G0/G1期进程,促进PTC细胞增殖并抑制凋亡。Objective To investigate the expression of apolipoprotein C1(APOC1)in papillary thyroid carcinoma(PTC)and its effects on proliferation and apoptosis of PTC cells.Methods The expression level of APOC1 in PTC and its impact on prognosis were analyzed using GEPIA 2 and Kaplan-Meier databases.Immunohistochemistry(IHC)and Western blotting were used to detect the expression of APOC1 in PTC and adjacent tissues and in 3 PTC cell lines and normal thyroid Nthyori 3-1 cells.In TPC-1 and BCPAP cells,the effect of Lipofectamine 2000-mediated transfection with APOC1 siRNA or an APOC1-overexpressing plasmid on cell growth and colony formation ability were examined by observing the growth curves and using colony-forming assay.The changes in cell cycle and apoptosis of the transfected cells were analyzed with flow cytometry.RT-qPCR and Western blotting were used to detect the changes in expressions of P21,P27,CDK4,cyclin D1,Bcl-2,Bax,caspase-3 and caspase-9 and the key proteins in the JAK2/STAT3 signaling pathway.Results APOC1 expression was significantly higher in PTC tissues and the 3 PTC cell lines than in the adjacent tissues and Nthyori 3-1 cells,respectively.In TPC-1 and BCPAP cells,APOC1 knockdown obviously reduced cell proliferative activity,increased the percentage of G0/G1 phase cells,lowered the percentages of S and G2 phase cells,promoted cell apoptosis,and downregulated mRNA and protein expression levels of CDK4,cyclin D1 and Bcl-2 and the protein levels of p-JAK2 and p-STAT3.APOC1 overexpression in the cells produced the opposite effects on cell proliferation,apoptosis,cell cycle and the mRNA and protein expressions.The application of AG490,a JAK2 inhibitor,strongly attenuated APOC1 overexpression-induced activation of the JAK2/STAT3 signaling pathway in BCPAP cells.Conclusion APOC1 overexpression promotes proliferation and inhibits apoptosis of PTC cells possibly by activating the JAK2/STAT3 signaling pathway and accelerating cell cycle progression.

关 键 词：甲状腺乳头状癌 载脂蛋白C1 增殖 凋亡 JAK2/STAT3信号通路 

分 类 号：R736.1[医药卫生—肿瘤]