肝癌阿帕替尼耐药细胞株的构建及耐药基因研究  

Construction of apatinib-resistant cell lines in hepatocellular carcinoma and study of resistant genes

作　　者：博伦 王祥旭 纪洪辰 艾丽萍 白引苗 刘洋 张红梅 BO Lun;WANG Xiangxu;JI Hongchen;AI Liping;BAI Yinmiao;LIU Yang;ZHANG Hongmei(Department of Clinical Oncology,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;The 965th Hospital of the Joint Logistics Support Force of the People’s Liberation Army of China,Jilin 132011,China)

机构地区：[1]空军军医大学西京医院肿瘤科,陕西西安710032 [2]解放军联勤保障部队第九六五医院,吉林吉林132011

出　　处：《空军军医大学学报》2024年第7期763-768,共6页Journal of Air Force Medical University

基　　金：陕西省自然科学基础研究计划重点项目(2021JZ-35);空军军医大学西京医院临床研究专项重点项目(XJZT24LZ15)。

摘　　要：目的探究肝癌细胞对阿帕替尼的潜在耐药机制。方法选取人SNU-739/SNU-878肝癌细胞系,构建阿帕替尼耐药细胞株。通过细胞活力检测和平板克隆形成实验对亲本及耐药细胞进行药敏试验,利用RNA芯片技术检测亲本及耐药细胞基因表达谱,根据表达差异筛选出介导阿帕替尼耐药的关键基因;利用qRT-PCR和Western blotting对关键基因进行验证,并通过药敏试验检测关键基因稳定过表达对细胞耐药的影响。结果阿帕替尼耐药细胞株的细胞活力、克隆形成数显著高于亲本细胞(P<0.05);筛选出关键基因氨基甲酰磷酸合成酶1(CPS1),其mRNA及蛋白水平在耐药细胞株中显著下调(P<0.05),过表达CPS1可显著降低肝癌细胞活力及克隆形成能力(P<0.05)。结论CPS1可能是介导肝癌细胞对阿帕替尼耐药的关键基因。Objective To explore the potential resistance mechanism of hepatocellular carcinoma cells to apatinib.Methods Human SNU-739/SNU-878 hepatocellular carcinoma cell lines were selected to construct apatinib-resistant cell lines.Antimicrobial susceptibility testing was carried out on parental and drug-resistant cells by cell viability detection and plate clone formation assay,the gene expression profiles of parental and drug-resistant cells were detected by RNA chip technology,and the key genes mediating apatinib resistance were screened out according to the expression differences.qRT-PCR and Western blotting were used to verify the key genes,and the effect of stable overexpression of key genes on cell drug resistance was detected by antimicrobial susceptibility testing.Results The cell viability and number of clones formed in apatinib-resistant cell lines were significantly higher than those in parental cells(P<0.05).A key gene,carbamoyl phosphate synthetase 1(CPS1),was screened out,and its mRNA and protein levels were significantly down-regulated in drug-resistant cell lines(P<0.05).Overexpression of CPS1 significantly reduced the viability and clonal formation ability of hepatocellular carcinoma cells(P<0.05).Conclusion CPS1 may be a key gene that mediates apatinib resistance in hepatocellular carcinoma cells.

关 键 词：肝癌 阿帕替尼 耐药 氨基甲酰磷酸合成酶1 

分 类 号：R735.7[医药卫生—肿瘤]