干涉HOTAIR对HepG2肝癌细胞增殖和周期的影响

Effects of siHOTAIR on proliferation and cell cycle of liver cancer cell HepG2

目的探讨干涉HOX转录物反义RNA(HOX transcript antisense RNA,HOTAIR)对HepG2肝癌细胞增殖和周期的影响。方法以人正常肝细胞系Chang liver为对照,采用RT-PCR分析HepG2、SMMC-7721、Bel-7402和MHCC97肝癌细胞中HOTAIR的表达水平,以HOTAIR表达显著增高的肝癌细胞系为模型,分两组进行实验,阴性对照组(NC组)转染阴性对照siRNA,干涉HOTAIR组(siHOTAIR组)分别转染3种siHOTAIR(siRNA#1、siRNA#2、siRNA#3),通过RT-PCR确定干涉效果最显著的siHOTAIR,然后MTT检测细胞增殖能力的变化,流式细胞术检测细胞周期的改变。结果与Chang liver相比,HepG2、SMMC-7721、Bel-7402和MHCC97细胞中HOTAIR的表达均增高,以HepG2增高最显著(P<0.01)。与NC组比较,siHOTAIR组中siRNA#2的干涉效果最明显(P<0.01),且siHOTAIR组(siRNA#2)HepG2细胞增殖能力显著降低,并发生G1期阻滞(P<0.01)。结论干涉HOTAIR能够抑制HepG2细胞增殖并诱导G1期阻滞。

Objective To elucidate the effects of interference of HOX transcript antisense RNA(HOTAIR)on proliferation and cell cycle of liver cancer cell HepG2.Methods With Chang liver cell line as the control,the expression levels of HOTAIR in liver cancer cell lines HepG2,SMMC-7721,Bel-7402 and MHCC97 were analyzed by RT-PCR.The cell line with the highest expression level of HOTAIR was chosen for the further experiment.The cells were transfected with NC siRNA in negative group and one of three siRNAs targeting HOTAIR(siRNA#1,siRNA#2 and siRNA#3)in siHOTAIR group,respectively.The most effective siHOTAIR was determined for transfection by RT-PCR.Cell proliferation ability was detected by MTT assay.Cell cycle was analyzed by flow cytometry.Results The expression level of HOTAIR was higher in HepG2,SMMC-7721,Bel-7402 and MHCC97 cell lines than that in normal Chang liver cell line,especially in HepG2 cells(P<0.01).Compared wtih NC group,the expression of HOTAIR in HepG2 cells was significantly downregulated in siHOTAIR groups,and the efficacy was the best after transfection with siRNA#2(P<0.01).HepG2 cell proliferation was inhibited and the cell cycle was arrested in G1 phase by siHOTAIR(siRNA#2)(P<0.01).Conclusion Interference of HOTAIR can inhibit HepG2 cell proliferation and induce G1 phase arrest.