苯乙酸对肝癌细胞系SMMC-7721的增殖抑制作用及与RNA编辑酶ADAR1表达的相关性

Inhibitory Effects of Phenylacetate on Proliferation and RNA Editing Deaminase ADAR1 Expression of Hepatocellular Carcinoma Cells SMMC-7721

为探讨苯乙酸(PA)对肝癌细胞系SMMC-7721的增殖抑制作用及其与RNA编辑酶ADAR1表达的相关性,应用细胞计数及MTT法检测了不同浓度(0.5,1.0,2.0和4.0 mmol/L)PA对肝癌细胞系SMMC-7721的增殖抑制作用,通过流式细胞术(FCM)分析了各细胞周期的细胞百分比,应用半定量逆转录-聚合酶链式反应(RT-PCR)及免疫印迹杂交分析使用不同浓度(0.5,1.0,2.0 mmol/L)PA作用后肝癌细胞系SMMC-7721中RNA编辑酶ADAR1 mRNA及蛋白表达的变化.结果表明,肝癌细胞系SMMC-7721经不同浓度PA作用后,增殖抑制率随作用时间延长及PA浓度增加而明显提高(P<0.05),但2.0和4.0 mmol/L PA作用72 h后组间差异比较无统计学意义(P>0.05).肝癌细胞系SMMC-7721中RNA编辑酶ADAR1 mRNA及蛋白表达随PA浓度增加而明显降低(P<0.05).通过沉默SMMC-7721细胞中ADAR1的表达发现,ADAR1表达下调可有效抑制肝癌细胞增殖.结果表明,PA可阻抑肝癌细胞系SMMC-7721细胞增殖,且存在时间及剂量的依赖性,作用机制与PA下调ADAR1表达相关.

To evaluate the inhibitory effects of phenylacetate （PA） on proliferation and RNA editing deami- nase ADAR1 expression of hepatocellular carcinoma cells SMMC-7721, the inhibitory effects of different con- centrations of PA（0. 5, 1.0, 2. 0 and 4. 0 mmol/L） on hepatocellular carcinoma cells SMMC-7721 were de- tected by MTT and cell numeration methods. The cell percentages were tested by flow cytometry（FCM）. The inhibitory effects of different doses of PA（0. 5, 1.0 and 2.0 mmol/L） on the expression of RNA editing dea- minase（ ADAR1 mRNA and protein） in SMMC-7721 cells were evaluated by RT-PCR and Western Blotting. With the increasing of concentrations of PA from 0. 5 to 4. 0 mmol/L and the prolonging time from 24 h to 72 h, the inhibitory rates of proliferation were significantly increased （ P 〈 0. 05 ） , but there was no significant difference between 2. 0 and 4.0 mmol/L PA group for 72 h（P〉0.05 ）. With the increasing of concentrations of PA, the expression of RNA editing deaminase （ADAR1 mRNA and protein） was decreased （P〈0. 05 ）. Fur- ther, we made down-regulation of the expression of ADAR1, the results showed that cancer cell proliferation was effectively inhibited. PA could inhibit the proliferation of hepatocellular carcinoma cells SMMC-7721 by the down-regulation of the expression of ADAR1 mRNA and protein in dependence on the time and doses.