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研究12C6+离子辐照对体外培养人肝癌细胞SMMC-7721细胞周期和P53、MDM2及P21表达的影响。采用0、1.0、2.0、4.0、6.0Gy12C6+离子束辐照细胞,用克隆形成法观察细胞存活情况;同时在辐照24h后用流式细胞仪检测细胞周期的变化,Western-blot检测细胞中P53、MDM2及P21蛋白表达情况。结果发现,重离子辐照后细胞存活率显著下降;1.0Gy、4.0Gy和6.0Gy照射组发生G0/G1期阻滞,而2.0Gy照射组出现G2/M期阻滞;Western-blot结果显示细胞辐照后MDM2的57kD蛋白表达水平无明显变化,而76kD蛋白表达水平随辐照剂量逐渐上升;P53和P21蛋白表达水平随辐照剂量增高。以上结果提示不同剂量的12C6+离子束照射可激活SMMC-7721细胞不同的细胞周期检测点,其中G0/G1期阻滞与P53和P21蛋白以及MDM2截短体76kD蛋白的表达水平升高有关。
The effects of 12C6+ ion irradiation on cell cycle and expression of P53, MDM2 and P21 in human hepatocellular carcinoma cell line SMMC-7721 were investigated. The cells were irradiated with 0, 1.0, 2.0, 4.0, and 6.0Gy12C6+ ion beams. The cell survival was observed by cloning method. At the same time, the changes of cell cycle were detected by flow cytometry 24 h after irradiation, and the P53 in cells was detected by Western-blot. , MDM2 and P21 protein expression. The results showed that cell viability after heavy ion irradiation decreased significantly; G0/G1 phase arrest occurred in the 1.0 Gy, 4.0 Gy, and 6.0 Gy irradiation groups, and G2/M phase arrest occurred in the 2.0 Gy irradiation group; Western-blot results showed There was no significant change in the 57kD protein expression level of MDM2 cells after irradiation, while the 76kD protein expression level gradually increased with the irradiation dose; the expression levels of P53 and P21 protein increased with the irradiation dose. The above results suggest that different doses of 12C6+ ion beam irradiation can activate different cell cycle detection sites of SMMC-7721 cells, and G0/G1 phase arrest is related to the increased expression of P53 and P21 proteins and 76kD protein of MDM2 truncates.