丙戊酸钠调节组蛋白乙酰化抑制胃癌细胞增殖的实验研究  

Suppression of gastric carcinoma proliferation by regulating histone acetylizad levels with VPA

作　　者：赵兴海[1] 孙京杰[1] 刘爱武[2] 李凯[2] 曹莉莉[1] 时昌文[1] 

机构地区：[1]山东省千佛山医院普外中心实验室,山东济南250014 [2]山东省千佛山医院普外中心,山东济南250014

出　　处：《中华肿瘤防治杂志》2009年第8期587-591,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省医药卫生科技发展计划青年基金(2007QZ019)

摘　　要：目的:探讨应用组蛋白脱乙酰酶抑制剂丙戊酸钠(VPA)逆转染色体组蛋白低乙酰化水平对胃癌细胞增殖的影响及其分子机制。方法:应用0.75～4.0mmol/LVPA作用于胃癌细胞系BGC-823细胞后,MTT法检测细胞生长抑制,PI标记流式细胞术检测细胞周期,间接免疫荧光法分析Cyclin A、Cyclin D1、Cyclin E和p21Waf/cip1蛋白表达,RT-PCR检测分析Cyclin A、Cyclin D1、Cyclin E和p21Waf/cip1 mRNA表达。结果:经0.75～4.0mmol/LVPA作用24、48、72和96h,各实验组均出现了时间、剂量依赖趋势的生长抑制,生长抑制率为9%～66%,差异有统计学意义,P<0.001;对照组细胞增殖G1期所占比例为57.8%～60.9%;而实验组则随药物浓度、作用时间的不同而出现不同程度的细胞周期G1期阻滞,G1期比例为58.2%～79.4%,与对照组比较差异有统计学意义,P<0.001。VPA干预BGC-823细胞后,Cyc-lin A、Cyclin D1蛋白和mRNA表达明显下调而p21Waf/cip1蛋白和mRNA表达明显上调,与对照组比较差异均有统计学意义,P<0.001;Cyclin E蛋白和mRNA表达变化未见明显差异,P>0.05。结论:组蛋白特异性脱乙酰酶抑制剂VPA可明显抑制胃癌细胞增殖、阻滞细胞增殖周期于G1期;其阻滞胃癌细胞增殖的作用通过上调p21Waf/cip1 mRNA、蛋白表达,下调Cyclin D1、Cyclin A mRNA和蛋白表达分别和(或)协同实现。OBJECTIVE： To investigate the suppressing effect on gastric carcinoma proliferation by up-regulating histone aeetylizad level with a selective inhibitor of HDACs-valproate acid sodium （VPA） and the maehanism. METHODS： BGC-823 gastric carcinoma cells were cultured with 0.75 -- 4.0 mmol/L valproie acid （VPA）, and the inhibition rate was studied by MTT assay; and the cell cycle was analyzed by flow cytometry with PI assay. Meanwhile, the protein and mRNA expressions of Cyclin A, Cyclin D1, Cyclin E, and p21^wal/cipl were analyzed by indirect immunofluorescence technique and RT-PCR respectively. RESULTS： The inhibition rates of the experimental groups were from 9% to 66% which increased significantly （P〈 0. 001）, and the increasement depended on dose and action time. The percentages of GI phases in the cell cycle of the control groups remained the same and that were from 57. 8%--60. 9%. Contrast to the control groups, VPA induced a significant arrest in GI phase （P〈0. 001） and a total of 58. 2% -79. 4% of G1 phase were found, p21Waf/cipl was up-regulated both at the mRNA and protein levels, and Cyclin A and Cyelin D1 were down-regulated both at the mRNA and protein levels （P〈0. 001）. Conversely, The mRNA expression of Cyclin E was unchanged upon the treatment with VPA, nor did it show any difference in the protein level （P 〉0.05）. CONCLUSIONS： Up-regulating his tone acetylizad level by VPA can inhibit gastric carcinoma cell proliferation, induce cell cycle arrest in G1 phase. The mechanism underlying its effect on cell cycle arrest in G1 phase induetion includes to up-regulate p21^Waf/cipl mRNA and protein expressions and down-regulate Cyclin A, Cyclin D1 mRNA and protein expressions.

关 键 词：胃肿瘤 丙戊酸 细胞增殖 细胞系 肿瘤 

分 类 号：R735.2[医药卫生—肿瘤]