过表达LHPP对人胆管癌细胞增殖的作用及机制  

Effect of LHPP overexpression on cell proliferation of human cholangiocarcinoma cells and its mechanism

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作  者:宋春灼 李想 李俊俊 刘猛 付立跃 朱海涛 SONG Chunzhuo;LI Xiang;LI Junjun;LIU Meng;FU Liyue;ZHU Haitao(Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Clinical Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区:[1]贵州医科大学附属医院肝胆外科,贵州贵阳550004 [2]贵州医科大学临床医学院,贵州贵阳550004

出  处:《贵州医科大学学报》2021年第9期999-1004,1041,共7页Journal of Guizhou Medical University

基  金:国家自然科学基金(81960429)。

摘  要:目的探究过表达磷脂磷酸组氨酸无机焦磷酸磷酸酶(LHPP)对人胆管癌(CCA)细胞增殖的作用和机制。方法取正常肝内胆管上皮细胞株HIBEpiC和人CCA细胞株TFK-1、HuCCT-1于RPMI-1640培养基培养,采用Western blot检测3种细胞株中的LHPP表达;慢病毒分别感染TFK-1和HuCCT-1细胞后分为过表达LHPP组(LHPP组)及对照组(NC组),采用细胞增殖实验(CCK-8)检测2组TFK-1和HuCCT-1细胞分别于接种后0、24、48及72 h的吸光度(OD)值,采用细胞克隆实验检测2组TFK-1和HuCCT-1细胞的克隆数,采用Western blot分析2组TFK-1和HuCCT-1细胞中人蛋白激酶B(AKT)、人磷酸化蛋白激酶B(p-AKT)及人抑癌基因P53蛋白表达水平。结果人CCA细胞株TFK-1和HuCCT-1细胞中LHPP蛋白表达量较人正常肝内胆管上皮细胞HIBEpiC相对下降(P<0.05),LHPP组TFK-1和HuCCT-1细胞中LHPP蛋白表达量均较NC组增高(P<0.05);LHPP组TFK-1和HuCCT-1细胞于48 h和72 h时OD值均较NC组减少(P<0.05);LHPP组TFK-1和HuCCT-1细胞克隆形成能力较NC组降低(P<0.05);与NC组相比,LHPP组TFK-1和HuCCT-1细胞p-AKT蛋白表达下降、P53蛋白表达增高(P<0.05)。结论过表达LHPP能抑制CCA细胞的增殖,其机制可能与p-AKT表达下调、P53表达增加有关。Objective To explore the effect of overexpression of phospholysine phosphohistidine inorganic pyrophosphate phosphatase(LHPP)on the proliferation of human cholangiocarcinoma(CCA)cells and its mechanism.Methods Normal intrahepatic bile duct epithelial cell line HIBEpiC and human CCA cell lines(TFK-1 and HuCCT-1)were cultured in RPMI-1640 medium.Western blot was used to detect the expression of LHPP in these three cell lines.TFK-1and HuCCT-1 cells were transfected with expression vector carrying LHPP(LHPP group)or expression vector carrying negative control(NC group).Cell Counting Kit-8(CCK-8)assay was used to detect absorbance(OD)values of two groups of TFK-1 and HuCCT-1 cells at 24,48,and 72 h after seeding.Cologenic formation assay was used to measure colony numbers of two groups of TFK-1 and HuCCT-1 cells.The expression levels of Kinase B(AKT),phosphorylated AKT(p-AKT),and human tumor suppressor gene(P53)in two groups were detected using Western blot.Results LHPP expression levels in human CCA TFK-1 and HuCCT-1 cells were relatively lower than that in HIBEpiC(P<0.05).LHPP protein expression levels in LHPP groups of TFK-1 and HuCCT-1 cells were higher than that in NC groups(P<0.05).The OD values of TFK-1 and HuCCT-1 cells in LHPP groups were lower than those in NC group at 48 h and 72 h(P<0.05).Colony numbers TFK-1 and HuCCT-1 cells in LHPP groups were less than those in the NC group(P<0.05).When compared with the NC group,the expression levels of p-AKT protein in TFK-1 and HuCCT-1 cells in LHPP groups were decreased,but the expression level of P53 protein was increased(P<0.05).Conclusion Overexpression of LHPP can inhibit the proliferation of CCA cells,and its mechanism may be related to the down-regulation of p-AKT expression and the increase of P53 expression.

关 键 词:胆管肿瘤 细胞增殖 LHPP基因 慢病毒 蛋白激酶B 磷酸化蛋白激酶B 

分 类 号:R735.9[医药卫生—肿瘤]

 

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