CXC趋化因子配体10对肝细胞癌SMMC-7721细胞增殖和迁移的影响及其机制  

Effect of CXC chemokine ligand 10 on proliferation and migration of hepatocellular carcinoma SMMC-7721 cells and its mechanism

作　　者：邓文俊 胡连涛 赵彬男 董新宇 李学斌 李杰 杨馨妍 郭晓莉 李玥 曲义坤[5] 王伟群 DENG Wenjun;HU Liantao;ZHAO Binnan;DONG Xinyu;LI Xuebin;LI Jie;YANG Xinyan;GUO Xiaoli;LI Yue;QU Yikun;WANG Weiqun(Department of Physiology,School of Basic Medical Sciences,Jiamusi University,Jiamusi 154007,China;Microbial-Immunomodulatory Network and Related Diseases Key Laboratory,Heilongjiang Province,Jiamusi 154007,China;Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Jiamusi University,Jiamusi 154007,China;Department of Laboratory,First Affiliated Hospital,Jiamusi University,Jiamusi 154007,China;Department of General Surgery,First Affiliated Hospital,Jiamusi University,Jiamusi 154007,China)

机构地区：[1]佳木斯大学基础医学院生理学教研室,黑龙江佳木斯154007 [2]黑龙江省微生物-免疫调节网络与相关疾病重点实验室,黑龙江佳木斯154007 [3]佳木斯大学基础医学院生物化学与分子生物学教研室,黑龙江佳木斯154007 [4]佳木斯大学附属第一医院检验科,黑龙江佳木斯154007 [5]佳木斯大学附属第一医院普外科,黑龙江佳木斯154007

出　　处：《吉林大学学报（医学版）》2023年第5期1227-1233,共7页Journal of Jilin University：Medicine Edition

基　　金：黑龙江省科技厅自然科学基金联合引导项目(LH2021H109)。

摘　　要：目的:探讨外源性CXC趋化因子配体10 (CXCL10)对肝细胞癌(HCC) SMMC-7721细胞增殖和迁移的影响,并阐明其作用机制。方法:按照CXCL10作用浓度,将人HCC SMMC-7721细胞分为0 mg·L^(-1)CXCL10组、10 mg·L^(-1)CXCL10组和30 mg·L^(-1)CXCL10组。上述部分细胞给予细胞外调节蛋白激酶(ERK)抑制剂PD98059 (80μmol·L^(-1))后,将SMMC-7721细胞分为0 mg·L^(-1)CXCL10+PD98059组、 10mg·L^(-1)CXCL10+PD98059组和30mg·L^(-1)CXCL10+PD98059组。CCK-8法检测各组SMMC-7721细胞增殖率,EdU法检测各组SMMC-7721细胞中EdU阳性表达率,Transwell小室实验检测各组SMMC-7721细胞迁移率,Western blotting法检测各组SMMC-7721细胞中ERK、磷酸化ERK (p-ERK)和细胞周期蛋白D1 (Cyclin D1)蛋白表达水平。结果:CCK-8法检测,培养24h后,与0mg·L^(-1)CXCL10组比较,10mg·L^(-1)CXCL10和30mg·L^(-1)CXCL10组SMMC-7721细胞增殖率升高(P<0.05或P<0.01)。EdU法检测,与0mg·L^(-1)CXCL10组比较,10mg·L^(-1)CXCL10和30mg·L^(-1)CXCL10组SMMC-7721细胞中EdU阳性表达率升高(P<0.01);Transwell小室实验检测,培养48 h后,与0 mg·L^(-1)CXCL10组比较,10 mg·L^(-1)CXCL10和30 mg·L^(-1)CXCL10组SMMC-7721细胞迁移率升高(P<0.01)。Western blotting法检测,细胞培养24 h后,采用CXCL10溶液处理24h,与0mg·L^(-1)CXCL10组比较,10mg·L^(-1)CXCL10组和30 mg·L^(-1)CXCL10组SMMC-7721细胞中ERK、p-ERK及Cyclin D1蛋白表达水平升高(P<0.01)。CCK-8法检测,加入ERK抑制剂PD98059后,与0 mg·L^(-1)CXCL10组比较,10 mg·L^(-1)CXCL10+PD98059组和30 mg·L^(-1)CXCL10+PD98059组SMMC-7721细胞增殖率降低(P<0.05);与10 mg·L^(-1)CXCL10组比较,10 mg·L^(-1)CXCL10+PD98059组SMMC-7721细胞增殖率降低(P<0.05);与30 mg·L^(-1)CXCL10组比较,30 mg·L^(-1)CXCL10+PD98059组SMMC-7721细胞增殖率降低(P<0.05)。结论:CXCL10能够促进HCCSMMC-7721细胞增殖和迁移,其作用机制与上调ERK/p-ERK/Cyclin D1通路蛋白表达有关。Objective:To discuss the effect of exogenous CXC chemokine ligand 10(CXCL10)on the proliferation and migration of the hepatocellular carcinoma(HCC)SMMC-7721 cells,and to clarify its mechanism.Methods:The human HCC SMMC-7721 cells were divided into 0 mg·L^(-1) CXCL10 group,10 mg·L^(-1) CXCL10 group,and 30 mg·L^(-1) CXCL10 group according to the CXCL10 concentration.Some of the above cells were treated with extracellular regulated protein kinase(ERK)inhibitor PD98059(80μmol·L^(-1)),then the SMMC-7721 cells were divided into 0 mg·L^(-1) CXCL10+PD98059 group,10 mg·L^(-1) CXCL10+PD98059 group,and 30 mg·L^(-1) CXCL10+PD98059 group.The proliferation rates of the SMMC-7721 cells in various groups were detected by CCK-8 method;the EdU positive expression rates in SMMC-7721 cells in various groups were detected by EdU method;the migration rates of the SMMC-7721 cells in various groups were detected by Transwell chamber assay;the expression levels of ERK,phosphorylated ERK(p-ERK),and Cyclin D1 proteins in the SMMC-7721 cells in various groups were detected by Western blotting method.Results:The CCK-8 results showed that after cultured for 24 h,compared with 0 mg·L^(-1) CXCL10 group,the proliferation rates of the cells in 10 mg·L^(-1) CXCL10 group and 30 mg·L^(-1) CXCL10 group were increased(P<0.05 or P<0.01);the EdU detection results showed that compared with 0 mg·L^(-1) CXCL10 group,the positive expression rates of EdU in the cells in 10 mg·L^(-1) CXCL10 group and 30 mg·L^(-1) CXCL10 group were increased(P<0.01).The Transwell chamber assay results showed that after cultured for 48 h,compared with 0 mg·L^(-1) CXCL10 group,the migration rates of the cells in 10 mg·L^(-1) CXCL10 group and 30 mg·L^(-1) CXCL10 group were increased(P<0.01).The Western blotting results showed that after cultured for 24 h and treated with CXCL10 for 24 h,compared with 0 mg·L^(-1) CXCL10 group,the expression levels of ERK,p-ERK,and Cyclin D1 proteins in the SMMC-7721 cells in 10 mg·L^(-1) CXCL10 group and 30 mg·L^(-1) CXCL10 gro

关 键 词：CXC趋化因子配体10 肝细胞肿瘤 细胞外调节蛋白激酶 周期蛋白D1 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]