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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 deaminase ADAR1 expression of hepatocellular carcinoma cells SMMC-7721, the inhibitory effects of different concentrations of PA(0.5, 1.0, 2.0 and 4.0 mmol/L) on hepatocellular carcinoma cells SMMC-7721 were detected 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). Further, 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.
