HDAC抑制剂SAHA对人肝癌细胞分化诱导作用的研究  

Effects of SAHA on the differentiation of HCC SMMC-7721 cells

作　　者：高山[1] 雷道雄[2] 艾中立[2] 魏芸[3] 

机构地区：[1]杭州市萧山区第一人民医院普外科,311201 [2]武汉大学中南医院普外科 [3]武汉大学医学院病毒学研究所

出　　处：《中华普通外科杂志》2009年第10期802-805,共4页Chinese Journal of General Surgery

基　　金：湖北省科技攻关项目（2001AA308B14） 志谢 感谢美国Aton Phanna公司惠赠SAHA试剂

摘　　要：目的探讨组蛋白去乙酰化酶（histone deacetylases，HDAC）抑制剂SAHA（suberoylanilide hydroxamic acid）对人肝癌SMMC-7721细胞的分化诱导作用。方法倒置显微镜观察SAHA对SMMC-7721细胞形态的影响；MTT比色法测定SAHA对SMMC-7721细胞增殖的抑制情况；免疫细胞化学检测SAHA对SMMC-7721细胞中甲胎蛋白（AFP）和增殖细胞核抗原（PCNA）表达的影响；流式细胞术（FCM）分析细胞周期；RT—PCR方法检测处理前后SMMC-7721细胞p21WAF1基因mRNA的表达变化。结果实验组细胞增殖速度显著减慢，与正常细胞形态变化相似；MTT比色法测定结果显示不同浓度SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0／G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0／G1期；RT—PCR检测结果表明，实验组细胞中p21WAF1 mRNA的表达明显增加。结论SAHA对人肝癌细胞具有显著的诱导分化作用，诱导肝癌细胞分化的机理可能与抑制HDAC的活性，上调p21WAF1 mRNA表达，及阻滞肝癌细胞G0／G1期有关。Objective To investigate the effects of SAHA on the differentiation of human hepatocellular carcinoma （HCC） SMMC-7721 cells. Methods Cell morphology was examined by light microscopy; Cell viability was determined by MTT assay; The expression of Alpha-fetoprotein （AFP） and proliferating cell nuclear antigen （PCNA） was analyzed with immunocytochemistry; Flow cytometry （FCM） was used to investigate the cell cycle; The expression of p21WAF1mRNA was detected by semi-quantitative RT-PCR. Results Light microscopy showed that SMMC-7721 cells induced by SAHA underwent restorational alterations in morphology which were different from those of nontreated cells but were similar to those of normal cells; MTT assay showed that SAHA inhibited the proliferation of SMMC-7721 cells in a dose-dependent and time-dependent way ; Immunocytochemistry assay showed that the expression of AFP and PCNA decreased significantly; FCM analysis showed SAHA could arrest SMMC-7721 cells at G0/G1 phase, with an accumulation of the cells at G0/G1 phase while a decrease of cells at S phase; Semi-quantitative RT-PCR detection revealed that the expression of p21WAF1 mRNA was upregulated remarkably in the cells treated with SAHA. Conclusion SAHA could induce differentiation of human HCC SMMC-7721 cells inhibiting enzymatic activity of HDAC, upregulating the expression of p21WAF1mRNA as well as causing arrest of HCC cells at G0/G1 phase.

关 键 词：肝肿瘤 细胞分化 细胞周期 基因表达调控 酶学 

分 类 号：R686[医药卫生—骨科学]