摘要
目的探究microRNA-122(miR-122)对肝肿瘤细胞HepG2增殖的抑制效应。方法培养肝肿瘤细胞HepG2并随机分为miR-122组、阴性对照组(NC组)、空白对照组,miR-122组使用LipofectamineTM2000试剂转染肝癌细胞株HepG2细胞,NC组将阴性对照siRNA转染HepG2细胞,空白对照组只加胎牛血清RPMI-1640培养基。转染24 h后,采用MTS试剂盒测定细胞活力,计算细胞增殖抑制率,采用荧光定量PCR测定细胞中凋亡抑制蛋白(XIAP)、细胞周期素D1(Cyclin D1)、细胞周期蛋白依赖激酶2(CDK2)、细胞周期蛋白依赖激酶4(CDK4)、血管内皮生长因子(VEGF)、VEGFR1、VEGFR2、肝细胞生长因子(HGF)的mRNA含量。结果干预后24 h,miR-122组细胞的吸光值(OD)明显低于空白对照组、NC组,细胞增殖抑制率明显高于NC组(t=9.43、19.475、P<0.05),miR-122组细胞中XIAP、Cyclin D1、CDK2、CDK4 mRNA含量明显低于空白对照组、NC组(t=8.135、7.542、11.740、5.937,P<0.05),VEGF、VEGFR1、VEGFR2、HGF mRNA含量明显低于空白对照组、NC组(t=7.741、13.002、6.347、6.662,P<0.05)。结论 miR-122能够抑制肝肿瘤细胞HepG2的增殖,靶向抑制细胞周期相关分子、血管新生相关分子的表达是miR-122发挥增殖抑制效应的可能机制。
Objective To investigate the inhibiting effect of microRNA-122（miR-122） on the proliferation of hepa- toeellular carcinoma cells HepG2. Methods The hepatocellular carcinoma cells HepG2 were cultured and randomly di- vided into miR-122 group, negative control group （NC group） and blank control group. In miR-122 group, the HepG2 cells were transfected by LipofectamineTM2000 reagent. In NC group, the HepG2 cells were transfected by siRNA. In blank control group, only the fetal bovine serum culture medium RPMI-1640 was added. Twenty-four hours after the transfection, the cell viability was measured by MTS kit, and the cellular proliferation inhibition rate was calculated. Fluorescence quantitative PCR was adopted to measure the mRNA levels of X linked inhibitor of apoptosis protein （XI- AP） , CyclinD1 , cyclin dependent kinase 2 （CDK2） , cyclin dependent kinase 4 （CDK4） , vascular endothelial growth factor （VEGF） , VEGFR1, VEGFR2 and hepatocyte growth factor （HGF）. Results Twenty-four hours after the treat- ment, the optical density （OD） of ceils in miR-122 group was significantly lower than that in blank control group and NC group, and the cellular proliferation inhibition rate was significantly higher than that in the NC group （t = 9.43, 19. 475, P 〈 0.05）. The mRNA levels of XIAP, CyclinD, CDK2 and CDK 4 in miR-122 group were significantly low- er than those in blank control group and NC group （t =8. 135, 7. 542, 11. 740, 5. 937, P 〈0.05） , and the mRNA levels of VEGF, VEGFR1, VEGFR2 and HGF were significantly lower than those in blank control group and NC group （t = 7. 741, 13. 002, 6. 347, 6. 662, P 〈 0.05）. Conclusion microRNA-122 can inhibit the proliferation of hepato- cellular carcinoma cell HepG2, and the possible mechanism is targeted inhibition of the expression of cell cycle relevant molecules and angiogenesis relevant molecules.
出处
《胃肠病学和肝病学杂志》
CAS
2016年第12期1467-1470,共4页
Chinese Journal of Gastroenterology and Hepatology
