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目的研究低剂量辐射(LDR)对人肝癌细胞(BEL7402)增殖、细胞周期及相关信号蛋白表达的影响,为LDR临床应用提供理论依据。方法体外培养BEL7402,分为7个实验组:假辐射组(0 mGy)、5个LDR组(25,50,75,100,200 mGy)及高剂量辐射组(1 Gy)。辐射后用细胞计数法及噻唑蓝比色试验(MTT)检测细胞增殖率,用流式细胞术测定细胞周期中G0/G1,S,G2/M期的比例。应用蛋白分析系统分析检测细胞周期及相关信号蛋白共40个蛋白的表达。结果细胞计数及MTT试验,5个LDR组BEL7402细胞增殖率与假辐射组比较差异不显著(P>0.05)。流式细胞术试验,5个LDR组BEL7402细胞12、24 h G0/G1期细胞比例增高,但与假辐射组比较差异不显著(P>0.05);S期比例降低,与假辐射组比较差异显著(P<0.05);G2/M期比例升高,与假辐射组比较差异显著(P<0.05)。48 h后G0/G1,S,G2/M期与假辐射组比较差异不显著(P>0.05)。蛋白分析试验LDR后8种蛋白表达降低。结论LDR对BEL7402细胞增殖无促进作用,但在一定程度抑制增殖及细胞周期相关蛋白的表达。
Objective To study the effects of low dose radiation (LDR) on the proliferation, cell cycle and related signaling proteins in human hepatocellular carcinoma (BEL7402) cells, and to provide a theoretical basis for the clinical application of LDR. Methods BEL7402 was cultured in vitro and divided into seven experimental groups: sham irradiation group (0 mGy), 5 LDR group (25,50,75,100,200 mGy) and high dose radiation group (1 Gy). Cell proliferation rate was measured by cytometry and MTT assay after radiation. The proportion of G0 / G1, S, G2 / M phase in the cell cycle was determined by flow cytometry. Protein analysis system was used to detect the expression of 40 proteins in cell cycle and related signaling proteins. Results Cell count and MTT assay showed that the proliferation rate of BEL7402 cells in five LDR groups was not significantly different from that in sham irradiation group (P> 0.05). Flow cytometry showed that the percentage of cells in G0 / G1 phase of 5 LDR group was increased at 12 and 24 h, but there was no significant difference compared with that of sham radiation group (P> 0.05); the proportion of S phase was lower than that of sham radiation group (P <0.05). The proportion of G2 / M phase increased significantly compared with sham radiation group (P <0.05). After 48 h, the G0 / G1, S, G2 / M phases were not significantly different from the sham radiation group (P> 0.05). After protein analysis, the expression of eight proteins decreased after LDR test. Conclusion LDR has no effect on the proliferation of BEL7402 cells, but inhibits the proliferation and the expression of cell cycle related proteins to a certain extent.