摘要目的 比较3株肝癌耐药细胞(Huh-7/ADM、Huh-7/CBP、Huh-7/MMC)与亲本肝癌细胞Huh-7的形态学差异；分析这3株肝癌耐药细胞及Huh-7的miRNA表达谱,筛选3株耐药细胞中异常表达的微小RNA(miRNA).方法 光学显微镜下观察各细胞株细胞形态学特点、透射电镜下观察各细胞株细胞内超微结构特点.miRNA芯片检测各细胞株miRNA表达谱,real-time定量PCR(qPCR)法验证芯片结果.结果 镜下见肝癌耐药细胞比亲本肝癌细胞Huh-7更具多形性,核异型性亦增加；细胞周边微绒毛减少；胞质增多,胞质内细胞器含量增加,并出现脂滴、糖原等成分.miRNA表达谱分析显示:与Huh-7相比,有32个miRNA在3种肝癌耐药细胞株中表达同时上调,有22个miRNA在3种耐药细胞株中表达同时下调.real-time qPCR检测证实miR-15a、miR-16、miR-27b、miR-30b、miR-146a、miR-146b-5p、miR-181a、miR-181d、miR-194在各肝癌耐药细胞中表达同时上调.结论 耐药细胞异型性增加,筛选出的肝癌耐药细胞中异常表达miRNA,可为研究肝癌多药耐药与miRNA的相关性提供依据.

abstractsObjective To compare morphological differences of three drug-resistant hepatocellular carcinoma (HCC) cell subclones (Huh-7/ADM,Huh-7/CBP,Huh-7/MMC) and their parental Huh-7 cell line,to analyze differential microRNA(miRNA) expression profiles in these cells and,finally to screen for the abnormal expressed miRNAs in drug-resistant HCC cells.Methods Cellular morphology was observed by histology and transmission electron microscopy.MiRNA microarray was used to analyze the differential miRNA expression profiles in these cells (Huh-7,Huh-7/ADM,Huh-7/CBP,Huh-7/MMC) followed by real time quantitative PCR validation.Results The drug-resistant cells had more intracytoplasmic organelles and were larger in size along with increased cytological pleomorphism than the parental Huh-7 cells.Compared with the parental Huh-7 cells,32 simultaneously up-regulated and 22 down-regulated miRNAs were found in three drug-resistant cells.Up-regulation of miR-15a,miR-16,miR-27b,miR-3Ob,miR-146a,miR-146b-5p,miR-181a,miR-181d and miR-194 was verified by RT-qPCR.Conclusion Drug-resistant HCC cells have abnormal expressed miRNAs,which may be explored to further investigate the association of miRNA expressions with multidrugs resistance in HCC.