摘要
目的观察RNA干扰下调ADAR1基因表达对肝癌细胞增殖能力的影响。方法用小分子干扰RNA(siRNA)转染肝癌细胞SMMC-7721,RT-PCR法检测ADAR1mRNA表达,Westernblotting法测定ADAR1蛋白,MTr比色法检测SMMC-7721细胞增殖。结果细胞转染24、48、72小时后,实验组ADAR1mRNA相对表达量分别为0.612±0.086、0.264±0.018、0.156±0.063,对照组分别为1.032±0.107、0.898±0.092、0.968±0.074,差异具有统计学意义(P〈0.05);对照组与空白组ADAR1mRNA表达差异无统计学意义(P〉0.05)。实验组ADAR1蛋白相对表达量分别为0.684±0.079、0.324±0.042、0.145±0.058,对照组分别为1.002±0.092、0.917±0.068、0.972±0.073,差异均有统计学意义(P〈0.05);对照组与空白组ADAR1mRNA表达差异无统计学意义(P〉0.05)。细胞转染后SMMC.7721细胞增殖活性明显下降(P〈0.05)。结论ADAR1基因特异性siRNA干扰后,ADAR1mRNA及蛋白相对表达水平显著下降,SMMC-7721细胞的增殖能力亦明显下降。
Objective To observe the impact of RNA interference-induced ADAR1 down-regulation on cell proliferation of liver cancer. Methods Small interfering RNA (siRNA) was transfected into liver cancer cell line SMMC-7721, ADAR1 expression was detected by RT-PCR and Western blotting. Cell prolif- eration was determined by methyl thiazol tetrazolium (MTF) assay. Results After transfection for 24, 48, and 72 h, ADAR1 mRNA expression was 0.612±0.086, 0.264±0.018, 0. 156±0.063 in experimental group and 1. 032±0. 107, 0. 898±0. 092, 0. 968±0. 074 in control group, respectively. Experimental group had significantly lower ADAR1 mRNA than the other groups (P 〈 0.05 ), and there was no statistically significant difference between control and blank group (P 〉 0. 05). ADAR1 protein relative expression was 0. 684±0. 079, 0. 324 ±0. 042, 0. 145±0. 058 in experimental group and 1. 002±0. 092, 0. 917± 0. 068, 0. 972±0.073 in control group, respectively, which was statistically significant ( P 〈 0.05 ). After transfeetion with siRNA, the proliferation ability of SMMC-7721 cells was enormously inhibited ( P 〈 0.05 ). Conclusion ADAR1 mRNA and protein level could be significantly decreased by specific RNA interfe- rence, and cell proliferation in SMMC-7721 ceils were also greatly inhibited.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2016年第7期482-484,共3页
Chinese Journal of Hepatobiliary Surgery