CD36与肝细胞癌细胞增殖和迁移的关系及其对人肝癌细胞异种移植裸鼠模型的影响  

Relationship between CD36 and cell proliferation and migration in hepatocellular carcinoma and its effect on human hepatocellular carcinoma cell xenograft models in nude mice

作　　者：张立洪 吴巍[1] 胥广才[1] 张培建[1] 陶立德[1] Zhang Lihong;Wu Wei;Xu Guangcai;Zhang Peijian;Tao Lide(Department of Hepatobiliary Surgery,Affiliated Hospital of Yangzhou University,Yangzhou 225012,China)

机构地区：[1]扬州大学附属医院肝胆外科,扬州225012

出　　处：《肿瘤研究与临床》2024年第2期98-104,共7页Cancer Research and Clinic

基　　金：国家自然科学基金(82001964)。

摘　　要：目的观察CD36在肝细胞癌组织和细胞株中的表达水平,探讨CD36对人肝细胞癌细胞株增殖、迁移能力及人肝癌细胞异种移植裸鼠模型的影响。方法基于癌症基因组图谱(TCGA)数据库相关信息分析371份肝细胞癌及癌旁组织中CD36转录本表达水平差异。前瞻性收集2019年1月至2021年2月就诊于扬州大学附属医院行手术治疗的48例肝细胞癌患者癌组织及相应的癌旁组织,采用实时荧光定量聚合酶链反应(qRT-PCR)法检测组织中CD36 mRNA水平。采用蛋白质印迹法检测人肝癌细胞株Huh7、HCCLM3及人正常肝细胞株LO2中CD36蛋白水平。将载有CD36干扰序列的质粒和空质粒转染到Huh7细胞或HCCLM3细胞,分别为sh-CD36组和对照组;采用CCK-8法检测培养0、12、24、36、48、60 h各组细胞的增殖能力(以吸光度值表示),采用划痕愈合实验、Transwell实验检测各组细胞迁移能力。将sh-CD36组或对照组Huh7细胞注射于BALB/c裸鼠腋窝皮下,每组4只,构建人肝癌异种移植裸鼠模型;接种1周后每周测量肿瘤长径、短径并计算肿瘤体积,接种5周后处死裸鼠,收集肿瘤标本并称量质量;显微镜下观察肿瘤组织细胞形态,采用免疫组织化学法检测肿瘤组织中CD36、Ki-67蛋白表达情况。结果对TCGA数据库数据分析显示,肝癌组织中CD36转录本水平较癌旁组织高(4.2±1.8比3.2±1.5,t=2.28,P=0.035)。qRT-PCR法对48例肝细胞癌患者组织检测显示,肝癌组织中CD36 mRNA相对表达量高于癌旁组织(0.76±0.26比0.48±0.23,t=3.52,P<0.001)。蛋白质印迹法检测显示,Huh7、HCCLM3细胞中CD36蛋白水平均高于LO2细胞,分别为LO2细胞的(1.42±0.11)倍和(1.68±0.16)倍(均P<0.001)。在mRNA及蛋白水平上,sh-CD36组Huh7和HCCLM3细胞CD36均低于对应的对照组(均P<0.001)。CCK-8法检测显示,sh-CD36组Huh7细胞和HCCLM3细胞分别于培养36 h和24 h开始增殖能力均低于对应的对照组(均P<0.01)。划痕愈合实验显示,培养48 h�Objective To observe the expression of CD36 in hepatocellular carcinoma tissues and cell lines,and to investigate the effects of CD36 on the proliferation and migration abilities of human hepatocellular carcinoma cell lines and human hepatocellular carcinoma cell xenograft models in nude mice.Methods Differences in the expression levels of CD36 transcripts in 371 hepatocellular carcinoma and paracancerous tissues were analyzed based on information from The Cancer Genome Atlas(TCGA)database.Cancer tissues and corresponding paracancerous tissues of 48 hepatocellular carcinoma patients who were diagnosed and underwent surgical treatment at the Affiliated Hospital of Yangzhou University from January 2019 to February 2021 were prospectively collected,and the levels of CD36 mRNA in the tissues were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)method.Western blotting was used to detect CD36 protein levels in human hepatocellular carcinoma cell lines Huh7 and HCCLM3 and human normal liver cell line LO2.Plasmids containing CD36 interfering sequences and empty plasmids were transfected into Huh7 cells or HCCLM3 cells for sh-CD36 group and control group,respectively.The CCK-8 assay was used to detect the proliferation ability(expressed as absorbance value)of cells in each group at 0,12,24,36,48 and 60 h of culture,and the scratch healing assay and Transwell assay were used to detect the migration ability of cells in each group.The Huh7 cells of sh-CD36 group or control group were injected into the axillary subcutis of BALB/c nude mice,with 4 mice in each group,to construct nude mice models of human hepatocellular carcinoma xenografts;the long and short diameters of tumor were measured weekly after 1 week of inoculation,and the tumor volume was calculated.The nude mice were put to death after 5 weeks of inoculation,and the tumor specimens were collected and weighed;the tumor cell morphology was observed under the microscope,and the expressions of CD36 and Ki-67 proteins in the tumor tissu

关 键 词：癌 肝细胞 抗原 CD36 细胞增殖 细胞运动 

分 类 号：R735.7[医药卫生—肿瘤]