缺氧诱导因子1α对肝癌细胞HepG2干细胞特性及表阿霉素敏感性的影响  

Effect of hypoxia-inducible factor-1αon stemness and epirubicin sensitivity of HepG2 hepatoma cells

作　　者：赵金金 张海光 崔非非 汪磊[1] 莫清江[1] 焦路阳[1] ZHAO Jinjin;ZHANG Haiguang;CUI Feifei;WANG Lei;MO Qingjiang;JIAO Luyang(Department of Clinical Laboratory,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang,Henan 453100,China;Department of Obstetrics and Gynecology,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang,Henan 453100,China)

机构地区：[1]新乡医学院第一附属医院检验科,河南新乡453100 [2]新乡医学院第一附属医院妇产科,河南新乡453100

出　　处：《临床肝胆病杂志》2021年第2期354-357,共4页Journal of Clinical Hepatology

基　　金：新乡医学院第一附属医院博士科研项目启动基金(10243);河南省医学科技攻关计划(联合共建)项目(LHGJ20190453)。

摘　　要：目的探讨缺氧诱导因子(HIF)1α对肝癌细胞HepG2干细胞特性及表阿霉素敏感性的影响。方法以肝癌细胞为研究对象,肝癌细胞HepG2脂质体转染过表达HIF-1α的质粒作为实验组,转染pcDNA3.1空质粒作为对照组,单独HepG2细胞为HepG2组。实时荧光定量PCR检测HIF-1αmRNA表达,Western Blot检测HIF-1α蛋白表达;流式细胞术检测细胞表面CD133表达。不同浓度表阿霉素(0、6.25、12.5、25、50μmol/L)作用3组细胞24 h,MTT法检测细胞活性,流式细胞术检测表阿霉素(50μmol/L)处理后细胞凋亡情况。计量资料多组间比较采用单因素方差分析,进一步两两比较采用t检验。结果相较于HepG2组及对照组,实验组HIF-1αmRNA的表达水平明显升高,差异具有统计学意义(P值均<0.001);Western Blot结果显示实验组HIF-1α蛋白高表达。HepG2组、对照组和实验组细胞的CD133比例分别为0.040%±0.003%、0.030%±0.010%、20.110%±0.600%,实验组的CD133阳性率显著高于HepG2组和对照组(P值均<0.001)。表阿霉素浓度为25、50μmol/L时,HepG2组和对照组的细胞活性明显受到抑制,显著低于实验组(P值均<0.05)。50μmol/L表阿霉素作用48 h后,实验组的细胞凋亡率(67.9%±2.5%)较HepG2组(93.6%±1.5%)和对照组(93.0%±1.2%)明显降低(P值均<0.001)。结论过表达HIF-1α的质粒成功转染至HepG2细胞,HIF-1α可提高肝癌细胞干细胞比例使其对表阿霉素耐药。Objective To investigate the effect of hypoxia-inducible factor-1α(HIF-1α)on the stemness and epirubicin sensitivity of hepatoma cells.Methods Hepatoma cells were selected for experiment.HepG2 hepatoma cells transfected with HIF-1αoverexpression plasmid were selected as experimental group,and those transfected with pcDNA3.1 empty plasmid were selected as control group;HepG2 cells alone were selected as HepG2 group.Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α;Western blot was used to measure the protein expression of HIF-1α;flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells.The three groups of cells were treated with epirubicin at different concentrations(0,6.25,12.5,25,and 50μmol/L)for 24 hours;MTT assay was used to measure cell viability,and flow cytometry was used to measure apoptosis after treatment with epirubicin(50μmol/L).A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the t-test was used for further comparison between two groups.Results Compared with the HepG2 group and the control group,the experimental group had a significant increase in the mRNA expression of HIF-1α(both P<0.001),and Western blot showed high expression of HIF-1αin the experimental group.The percentage of CD133 cells was 0.040%±0.003%in the HepG2 group,0.030%±0.010%in the control group,and 20.110%±0.600%in the experimental group,and the experimental group had a significantly higher positive rate of CD133+than the HepG2 group and the control group(both P<0.001).At an epirubicin concentration of 25 and 50μmol/L,the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group(both P<0.05).After the treatment with 50μmol/L epirubicin for 48 hours,the experimental group had a significantly lower cell apoptosis rate than the HepG2 group(67.9%±2.5%vs 93.6%±1.5%,P<0.001)and the control group(67.9%±2.5%vs 93.0%±1.2%,P<0.

关 键 词：肝肿瘤 实验性 缺氧诱导因子1 Α亚基 表柔比星 干细胞 抗药性 肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]