GCLM调控肝细胞肝癌细胞增殖及机制研究  

Study on the regulation and mechanism of GCLM in cell proliferation of hepatocellular carcinoma cells

作　　者：李茗鹤 白楠 孙笑 洪丹丹 李咏梅 LI Minghe;BAI Nan;SUN Xiao;HONG Dandan;LI Yongmei(Department of Pathogenic Biology,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China)

机构地区：[1]天津医科大学基础医学院病原生物学系,天津300070

出　　处：《天津医科大学学报》2024年第4期305-309,共5页Journal of Tianjin Medical University

基　　金：天津市教委科研计划项目(2022ZD055)。

摘　　要：目的:探讨γ-谷氨酰半胱氨酸合成酶修饰亚基(GCLM)对肝细胞肝癌(HCC)细胞增殖的作用及机制。方法:利用GEPIA、HPA数据库分析HCC组织和正常肝组织中GCLM表达情况;构建GCLM稳定敲降的HCC细胞株;CCK-8、克隆形成实验检测细胞增殖水平;铜测定实验检测细胞铜离子水平;铜死亡激活实验检测GCLM对细胞铜死亡的影响。结果:与正常肝组织相比,HCC组织中GCLM mRNA(P<0.0001)与蛋白表达水平明显升高;与对照组相比,GCLM敲降的MHCC97L细胞中GCLM的mRNA(F=255.20,P<0.001)与蛋白(F=50.77,P<0.01)表达水平下降;与对照组相比,GCLM敲降的MHCC97L细胞增殖减弱(F=42.79,P<0.01)、细胞克隆数量减少(F=1102,P<0.001);与对照组相比,GCLM敲降的MHCC97L细胞内铜离子水平升高(t=12.03,P<0.001);与对照组相比,Elesclomol和CuCl2联合处理的MHCC97L细胞增殖减弱(F=46.42,P<0.001),Elesclomol或CuCl2单独处理的MHCC97L细胞增殖均无明显变化(F=2.17,P=0.17)。结论:HCC组织中GCLM表达水平升高,GCLM敲降通过促进铜死亡来减弱HCC细胞增殖。Objective:To investigate the effect and underlying mechanism ofγ-glutamate-cysteine ligase modifier subunit(GCLM)on cell proliferation in hepatocellular carcinoma(HCC)cells.Methods:GEPIA and HPA databases were used to assess GCLM expression levels in HCC and normal liver tissues.HCC cells line with stable GCLM knockdown were constructed.The cell proliferation levels were detected by CCK-8,and colony formation assays.The level of Cu2+in cells was detected by copper assay.The effect of GCLM on cuproptosis was detected by the cuproptosis activation experiment.Results:Compared with normal liver tissues,GCLM mRNA(P<0.0001)and protein was highly expressed in HCC.Compared with the control group,the GCLM mRNA(F=255.20,P<0.001)and protein(F=50.77,P<0.01)expression levels were decreased in MHCC97L cells with GCLM knockdown.Compared with the control group,MHCC97L cells with GCLM knockdown showed decreased proliferation(F=42.79,P<0.01)and reduced colony formation ability(F=1102,P<0.001).Compared with the control cells,the Cu2+level of was significantly increased(t=12.03,P<0.001)in MHCC97L cells with GCLM knockdown.When combined treatment with Elesclomol and CuCl2,the proliferation of MHCC97L cells were inhibited(F=46.42,P<0.001)compared to the control cells.However,there was no significant effect on MHCC97L cells proliferation using Elesclomol or CuCl2 alone(F=2.17,P=0.17).Conclusion:GCLM is highly expressed in HCC tissues.GCLM knockdown inhibits HCC cell proliferation by promoting cell cuproptosis.

关 键 词：GCLM 肝细胞肝癌 铜死亡 

分 类 号：R735.7[医药卫生—肿瘤]