Dai Li-cheng, Wang Xiang, Yao Xing, Min Li-shan, Qian Fu-chu, He Jian-fang
Huzhou Key Laboratory of Molecular Medicine, Huzhou Central Hospital, Huzhou 313000, China.
Acta Pharmacol Sin. 2006 Nov;27(11):1453-8. doi: 10.1111/j.1745-7254.2006.00405.x.
To investigate the effect of antisense compounds (AS) targeting human p53 mRNA on radiosensitivity of MCF-7 cells.
Western blotting and RTPCR were used to analyze the protein content and mRNA level. Additionally, cell proliferation, cell cycle and cell apoptosis were all analyzed in irradiated or sham-irradiated cells.
Among the five antisense compounds (AS), AS3 was identified to efficiently inhibit p53 mRNA level and protein content. Interestingly, AS3 transfer has little effect on cell proliferation in DU-145 cells (mutant p53) after ionizing radiation (IR). In contrast, a marked increase of cell apoptosis and growth inhibition were observed in MCF-7 cells (wild-type p53), suggesting that AS3 can increase radiosensitivity of MCF-7 cells. Additionally, it was also observed that the transfection of AS3 decreased the fraction of G1 phase cells, and increased the proportion of S phase cells compared to untreated cells 24 h after IR in MCF-7 cell lines.
AS3 transfection increases MCF-7 cell apoptosis induced by 5 Gy-radiation, and this mechanism may be closely associated with abrogation of G1 phase arrest.
研究靶向人p53 mRNA的反义化合物（AS）对MCF-7细胞放射敏感性的影响。
采用蛋白质免疫印迹法和逆转录聚合酶链反应分析蛋白质含量和mRNA水平。此外，对辐照或假辐照的细胞进行细胞增殖、细胞周期和细胞凋亡分析。
在五种反义化合物（AS）中，AS3被确定能有效抑制p53 mRNA水平和蛋白质含量。有趣的是，电离辐射（IR）后，AS3转染对DU-145细胞（突变型p53）的细胞增殖影响不大。相反，在MCF-7细胞（野生型p53）中观察到细胞凋亡显著增加和生长抑制，表明AS3可提高MCF-7细胞的放射敏感性。此外，还观察到在MCF-7细胞系中，IR照射24小时后，与未处理细胞相比，AS3转染降低了G1期细胞比例，增加了S期细胞比例。
AS3转染增加了5 Gy辐射诱导的MCF-7细胞凋亡，这一机制可能与G1期阻滞的消除密切相关。
