LY294002调控JAK/STAT3通路对肝癌细胞增殖、凋亡的影响研究  

Effect of LY294002 regulating JAK/STAT3 pathway on proliferation and apoptosis of hepatocarcinoma cells

作　　者：蔡灵乐 谢毅[2] 张新明 王娜 CAI Ling-le;XIE Yi;ZHANG Xin-ming;WANG Na(Department of General Surgery,Ninth People's Hospital of Zhengzhou,Zhengzhou 450053,China;Department of Gastroenterology,Henan Provincial People's Hospital,Zhengzhou 450053,China)

机构地区：[1]郑州市第九人民医院普外科,河南郑州450053 [2]河南省人民医院胃肠外科,河南郑州450008

出　　处：《中国现代普通外科进展》2022年第5期337-342,共6页Chinese Journal of Current Advances in General Surgery

基　　金：2014年度河南省基础与前沿技术研究项目(142300410073)。

摘　　要：目的:探究LY294002调控Janus激酶(JAK)/信号传导及转录激活因子3(STAT3)通路对肝癌细胞增殖、凋亡的影响。方法:培养人肝癌SMMC-7721细胞至对数生长期后,分为对照组、LY294002处理组(加入10、20、30μmol/L)、白介素6处理组(IL-6组)、IL-6+LY294002处理组(IL-6+30μmol/L LY294002),干预培养48 h后,通过四甲基偶氮唑盐比色法(MTT)检测各组细胞活性,克隆形成实验检测各组细胞克隆形成能力,流式细胞术检测各组细胞凋亡情况,免疫印迹法(Western blot)检测各组细胞中Janus激酶2/信号传导及转录激活因子3(JAK2/STAT3)通路相关蛋白JAK2、磷酸化JAK2(p-JAK2)、STAT3、磷酸化STAT3(p-STAT3)及凋亡相关基因B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达。结果:与对照组相比,IL-6组细胞增殖抑制率、凋亡率、Bax蛋白表达显著降低(P<0.05),细胞克隆形成率、Bcl-2、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达显著升高(P<0.05);与IL-6组相比,10、20、30μmol/L LY294002组、IL-6+30μmol/L LY294002组细胞增殖抑制率、凋亡率、Bax蛋白表达显著升高(P<0.05),细胞克隆形成率、Bcl-2、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达显著降低(P<0.05);与10μmol/L、20μmol/L LY294002组相比,30μmol/L LY294002组细胞增殖抑制率、凋亡率、Bax蛋白表达显著升高(P<0.05),细胞克隆形成率、Bcl-2、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达显著降低(P<0.05),IL-6+30μmol/L LY294002组细胞增殖抑制率、凋亡率、Bax蛋白表达显著降低(P<0.05),细胞克隆形成率、Bcl-2、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达显著升高(P<0.05)。结论:LY294002可能通过调控JAK/STAT3通路抑制STAT3激活,从而抑制肝癌SMMC-7721细胞增殖,促进其凋亡。Objective:To investigate the effects of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PI3K/Akt)signal pathway blocker(LY294002)regulating Janus kinase(JAK)/signal transducers and activators of transcription 3(STAT3)pathway on the proliferation and apoptosis of hepatocarcinoma cells.Methods:Human hepatocarcinoma SMMC-7721 cells were cultured to logarithmic growth stage,and then divided into control group,LY294002 treatment group(adding 10,20,30μmol/L),interleukin 6 treatment group(IL-6 group),IL-6+LY294002 Treatment group(IL-6+30μmol/L LY294002),48 hours intervention culture,the cell activity of each group was detected by Methyl Thiazolyl Tetrazolium(MTT),the ability of cell clone formation was tested by clone formation experiment,the apoptosis was detected by flow cytometry,and Western blot was used to detect the expressions of the Janus kinase 2/signal transducers and activators of transcription 3(JAK2/STAT3)pathway related proteins JAK2,phosphorylated JAK2(p-JAK2),the apoptosis related gene B-cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 related X protein(Bax)were detected.Results:Compared with the control group,IL-6 group cell proliferation inhibition rate,apoptosis rate,Bax protein expression were significantly reduced(P<0.05),cell clone formation rate,Bcl-2,p-JAK2/JAK2,p-STAT3/STAT3 protein expression were significantly increased(P<0.05);compared with IL-6 group,10,20,30μmol/L LY294002 Group,IL-6+30μmol/L LY294002 group,cell proliferation inhibition rate,apoptosis rate,Bax protein expression increased significantly(P<0.05),cell clone formation rate,Bcl-2,p-JAK2/JAK2,p-STAT3/STAT3 protein expression was significantly reduced(P<0.05);compared with the 10μmol/L and 20μmol/L LY294002 groups,the cell proliferation inhibition rate,apoptosis rate,and Bax protein expression in the 30μmol/L LY294002 group were significantly increased(P<0.05),cell clone formation rate,Bcl-2,p-JAK2/JAK2,p-STAT3/STAT3 protein expression were significantly reduced(P<0.05),IL-6+30μmol/L LY294002 group cell proliferation inhibiti

关 键 词：LY294002 Janus激酶/信号传导及转录激活因子3通路 肝癌 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]