miRNA-221对肝癌HepG2细胞增殖及凋亡的影响  

Effect of miRNA-221 on proliferation and apoptosis of hepatocellular carcinoma HepG2

作　　者：韩双喜[1] 王丽[2] 王介营[1] 李传光[1] 张锋刚[1] 

机构地区：[1]滨州市中心医院普通外科,251700 [2]滨州市中心医院儿科,251700

出　　处：《国际医药卫生导报》2016年第8期1050-1054,共5页International Medicine and Health Guidance News

摘　　要：目的通过构建慢病毒介导的miRNA-221-siRNA，探讨其下调miRNA-221对肝癌HepG2细胞系生物学行为的影响。方法针对目标序列合成特异性siRNA干扰片段并克隆人慢病毒载体pGCSIL—GFP，将重组miRNA-221-RNAi—LV感染肝癌HepG2细胞系中，设为实验干扰组（miRNA-221-siRNA。SI组）、阴性对照组（negativecontrol，Nc组）和正常组（normal，N组）；分别采用聚合酶链反应方法检测miRNA-221目的基因的表达情况；用噻唑蓝比色法检测增殖情况、transwell小盒迁移实验检测迁移情况、Hochest33258凋亡实验检测调亡情况。结果（1）miRNA-221-RNAi—LV可显著降低miRNA-221的表达；（2）MTT实验96h，SI组吸光度显著低于NC组和N组（P=0．0032）；（3）迁移能力明显减弱（P=0．0004）；（4）SI组细胞凋亡明显增加，促凋亡基因Caspase3基因表达明显上调（P=0．0002）。结论miRNA-221-siRNA通过抑制miRNA-221的表达抑制肝癌HepG2细胞增殖，促进肝癌HepG2细胞凋亡，降低其迁移能力。Objective To study the effect of lentivirus-mediated miRNA-221 RNAi on the biological behaviors of human hepatic cancer cells. Methods miRNA-221 specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector pGCSIL-GFP. Hepatic cancer HepG2 cells were infected by microRNA-221-siRNA recombinant lentivirus（miRNA-221-siRNA-LV）, which were set as an SI group. The cells infected with miRNA-221-NC were set as an NC group. And the non-infected cells were set as an N group. The expression of the targeted genes of miRNA-221 were detected by transcription polymerase chain reaction （RT- PCR）. Cell proliferation was analyzed by MTT assay. Apoptosis was detected by Hoechst33258 and the apoptosis gene Caspase 3 was detect by RT-PCR. The invasion was detected by transwell method. BALB/c nude mice were subcutaneously transplanted with HepG2 cells. Results （1） miRNA-221-siRNA-LV could significantly lowered the expression of miRNA-221. （2） Hour 96 of MTT experiment, the absorbance was markedly lower in the SI group than in the NC group and the N group （P=-0.003 2）.（3） The number of cells that migrated through the chamber of the SI group was smaller than those of the NC group and the N group（P=0.000 4）.（4）The apoptosis was higher in the SI group than in the NC group and the N group. In addition, the expression of caspase 3 mRNA was significantly higher in the SI group than in the NC group and the N group （P=0.000 2）. Conclusions miRNA-221-siRNA suppresses the proliferation and migration of HepG2 cells and promote HepG2 cells＇ apoptosis by lowering the expression of miRNA-221.

关 键 词：miRNA-221 慢病毒 肝癌 HEPG2 

分 类 号：R735.7[医药卫生—肿瘤]