小分子干扰RNA下调星形细胞上调基因1表达对肝癌细胞增殖和凋亡的影响  

Effect of down-regulation of astrocyte elevated gene-1 expression by small interfering RNA inhibits cell proliferation and induces apoptosis in human hepatocellular carcinoma HepG2 cells

作　　者：王宏伟[1] 段长虹[2] 秦瑞英[3] 李伟伟[1] 侯丽娟[1] 段树鹏[1] 申保生[1] 郝洁[1] 杨玉新[4] 

机构地区：[1]新乡医学院第一附属医院感染疾病科,河南卫辉453100 [2]新乡医学院第一附属医院门诊部,河南卫辉453100 [3]新乡医学院第一附属医院妇产科,河南卫辉453100 [4]新乡医学院第一附属医院眼科,河南卫辉453100

出　　处：《中华实验外科杂志》2014年第5期982-984,共3页Chinese Journal of Experimental Surgery

基　　金：河南省教育厅资助项目（200733001）

摘　　要：目的探讨小分子干扰RNA（siRNA）下调星形细胞上调基因1（AEG-1）表达对肝癌细胞增殖和凋亡的影响及其机制。方法用设计合成的AEG-1siRNA转染人肝癌细胞HepG2细胞株，实时荧光定量逆转录聚合酶链反应（FQ—PCR）技术检测HepG2细胞AEG-1mRNA的表达；噻唑蓝（MTT）法检测HepG2细胞的增殖；流式细胞术检测AEG—lsiRNA对HepG2细胞凋亡和细胞周期的影响。结果AEG-1siRNA组AEG-1mRNA表达（0．43±0．04）明显低于空白对照组（0．96±0．09）和阴性对照组（0．94±0．08）（P〈0．01）；培养72h后AEG—1 siRNA组HepG2细胞吸光度（A）值（1．25±0．08）明显低于空白对照组（2．64±0．14）和阴性对照组（2．49±0．14，P〈0．01）；AEG-1s iRNA组HepG2细胞凋亡率（19．2±3．8）％明显高于空白对照组（5．3±1．4）％和阴性对照组（5．6±1．6）％（P〈0．01）；AEG-1 siRNA组HepG2细胞在DNA合成前期（G0／G1期）细胞比率[（61．38±2．31）％]明显高于空白对照组[（41．24±1．97）％]和阴性对照组[（42．15±1．64）％，P〈0．01]，而在合成期（S）[（25．36±1．21）％]及合成后期（G2／M）[（13．26±0．97）％]分别低于空白对照组[（33．60±1．24）％、（25．16±0．79）％]和阴性对照组[（32．52±1．23）％、（25．33±0．86）％，P〈0．01]。结论siRNA可通过下调HepG2细胞AEG-1基因的表达，抑制HepG2细胞的增殖，并促进其凋亡。Objective To investigate the effect of small interfering RNA （siRNA） -induced astro- cyte elevated gene-1 （ AEG-1 ） down-regnlation on the proliferation, apoptosis and cell cycle of human hepatocellular carcinoma cells and its mechanism. Methods siRNA and AEG-I siRNA were transfected to Human hepatocellular carcinoma HepG2 ceils, respectively. The expression of AEG-1 mRNA was detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction （FQ-PCR）. The cell proliferation was detected by methyl thiazol tetrazolium （MTF） method. The apoptosis and cell cycle were analyzed by flow cytometry. Results Expression of AEG-1 mRNA in AEG-1 siRNA group （0. 43 ± 0. 04） was significantly lower than that in control siRNA group （0. 94 ± 0. 08 ） and control group （ 0. 96 ± 0. 09 ） （P 〈0. 01）, respectively. After treated with AEG-1 siRNA for 72 h, the absorbance values of AEG-1 siRNA group （1. 25 ± 0. 08 ） was significantly lower than that in control siRNA group （2. 49 ± 0. 14） and control group （2. 64 ± 0. 14） （P 〈 0. 01 ）, respectively. The cell rate in G0/G1 phase （61.38 ± 2. 31 ）% was significantly higher than that in control siRNA group （42. 15 ± 1. 64）% and control group （41.24 ± 1.97）% （ P 〈 0. 01 ）, and the cell rates in S phase （ 25.36 ± 1.21 ） %, G2/M phase （ 13.26 ± 0. 97 ） % were lower than those in control siRNA group （ 32. 52 ± 1.23 ） %, （ 25.33 ± 0. 86 ） % and control group （ 33.60 ± 1.24）%, （25.16 ±0. 79）% （P 〈0. 01 ）, respectively. Conclusion The mRNA expression of AEG-1 is down-regulated by AEG-I siRNA in human hepatocellular carcinoma cells. Knockdown of AEG-1 expresion in human hepatocellular carcinoma cells significantly inhibits cell proliferation and apoptosis, and induces cell arrest in G0/G1 phase of the cell cycle.

关 键 词：星形细胞上调基因1 肝癌 小干扰RNA 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]