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目的:探讨苯丁酸钠诱导人肝癌细胞Bel7402,HepG2生长抑制、分化和细胞周期阻滞,以及对抑癌基因P21WAF1 /CIP1表达的影响。方法:培养Bel7402,HepG2,应用MTT比色法观察苯丁酸钠对Bel7402,HepG2的生长抑制作用,倒置相差显微镜观察细胞形态改变,流式细胞术分析细胞周期,以RTPCR检测Bel 7402,HepG2细胞中P21WAF1 /CIP1基因表达水平,WESTERN印记法检测P21WAF1 /CIP1蛋白的表达。结果:苯丁酸钠 2, 4, 8mmol·L-1处理 72h后Bel 7402, HepG2 的抑制率分别为 28. 43 %,57. 61 %, 71. 32 %和 27. 42 %, 57. 11 %, 70. 31 %处理后的细胞成纤维母细胞样改变,细胞阻滞于G期,P21WAF1 /CIP1基因和蛋白表达水平均增强。结论:苯丁酸钠抑制 2种人肝癌细胞株的生长,诱导部分人肝癌细胞分化,使细胞阻滞于G1期,苯丁酸钠能诱导P21WAF1 /CIP1基因的表达,增加P21WAF1 /CIP1蛋白的表达水平。
Objective: To investigate the effects of sodium phenylbutyrate on the growth inhibition, differentiation and cell cycle arrest of human hepatoma cells Bel7402 and HepG2, as well as the effect of P21WAF1/CIP1 on tumor suppressor gene expression. Methods: Bel7402 and HepG2 were cultured. The growth inhibitory effect of sodium phenylbutyrate on Bel7402 and HepG2 was observed by MTT colorimetric assay. Morphological changes were observed by inverted phase contrast microscope. The cell cycle was analyzed by flow cytometry. Bel 7402, HepG2 cells were detected by RTPCR. The expression level of P21WAF1/CIP1 gene was detected by Western blotting and the expression of P21WAF1/CIP1 protein was detected. Results: The inhibition rates of Bel 7402 and HepG2 after treatment with phenylbutyrate 2, 4, 8mmol·L-1 for 72h were 28.43%, 57. 61%, 71.32% and 27.42%, respectively, 57. 11 %, 70.31% of treated cells had fibroblast-like changes, cells were arrested in G phase, and the expression of P21WAF1/CIP1 gene and protein was enhanced. Conclusion: Phenylbutyrate inhibits the growth of two kinds of human hepatocellular carcinoma cell lines, induces the differentiation of some human hepatoma cells, and arrests cells in G1 phase. Phenylbutyrate induces the expression of P21WAF1/CIP1 gene and increases the expression of P21WAF1/CIP1 protein. The expression level.