Polo样激酶1小片段干扰RNA对肝癌细胞株HepG2的影响  

Effects of small interference RNAs targeting Polo-like kinase 1 in human liver cancer cell line HepG2

作　　者：张庆[1] 胡承明[1] 谭小平[2] 王卫政[2] 何长华[2] 刘南植[3] 

机构地区：[1]长江大学临床医学院内科教研室,湖北省荆州市434000 [2]长江大学附属第一医院消化内科,湖北省荆州市434000 [3]华中科技大学同济医学院附属同济医院消化内科,湖北省武汉市430030

出　　处：《世界华人消化杂志》2009年第24期2503-2507,共5页World Chinese Journal of Digestology

摘　　要：目的:联合RNA干扰技术探讨Polo样激酶1(PLK1)在肝癌的发生发展中的作用及对临床治疗的指导作用.方法:化学合成针对PLK1的小片段干扰RNA(PLK1-siRNA),转染至肝癌细胞株HepG2中,利用实时定量PCR、台盼蓝活性细胞计数和流式细胞术对PLK1的基因表达、细胞增殖、细胞周期和凋亡的检测.结果:转染PLK1-siRNA后,肝癌细胞中PLK1mRNA表达明显下降(P<0.01),表达量相当于空白组的49.7%±3.6%;细胞增殖活性从转染24h后明显下降;细胞分裂周期发生变化,S期比例下降,阻滞在G2/M期的细胞比例上升,于48、72h阻滞在G2/M期的细胞比例和细胞凋亡率与空白组相比均有显著增加(均P<0.05).结论:PLK1在肝癌的发生发展中起着重要作用,其siRNA能特异性抑制肝癌细胞中PLK1基因的表达,抑制细胞增殖,促进细胞凋亡,针对PLK1基因的RNA干扰有望用于肿瘤的基因治疗.AIM： To explore the role of PLK1 in carcinogenesis and development of liver cancer and its clinical significance for cancer treatment with the technique of RNA interference （RNAi）. METHODS： HepG2 cells were transfected with small interfering RNAs （siRNAs） targeting against the human PLK1 by chemosynthesis. The PLK1 mRNA levels of the cells transfected with siRNAs were monitored with real time PCR. Cell proliferation was evaluated by direct cell counting with trypan blue staining. Cell cycle and apoptosis were examined by flow cy- tometry. RESULTS： The PLK1 mRNA levels of trans- fected liver cancer cells were greatly lowered as compared with that of the control groups in 48 h after the siRNAs transfection （49.7% ＋ 3.6%）, （P 〈 0.01）. Cell proliferation was reduced from 24 h after transfection. Cell cycle distribution was changed and showed a strong G2/M arrest in 48 h and 72 h after transfection, and cell apoptosis was increased. There was significant difference as compared with the control group （both P 〈 0.05）. CONCLUSION： PLK1 plays an important role in the carcinogenesis and development of liver cancer, siRNAs targeting against human PLK1 may specially suppress the expression of PLK1 mRNA in liver cancer cells, inhibit cell prolif- eration and enhance cell apoptosis. The RNAi targeting PLK1 is expected to be used in gene therapy for human liver cancers.

关 键 词：POLO样激酶1 RNA干扰 实时定量聚合酶链式反应 肝癌 

分 类 号：R735.7[医药卫生—肿瘤] R557.1[医药卫生—临床医学]