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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 effect and mechanism of reversing the hypoacetylation of histone deacetylase (VPA) on the proliferation of gastric cancer cells using VPA. Methods: After treated with 0.75-4.0 mmol / L of LVPA for 24 hours, the growth of BGC-823 cells was detected by MTT assay. The cell cycle was detected by PI-flow cytometry. The expressions of Cyclin A, Cyclin D1, Cyclin E And p21Waf / cip1 protein expression, RT-PCR detection of Cyclin A, Cyclin D1, Cyclin E and p21Waf / cip1 mRNA expression. Results: After treated with 0.75-4.0 mmol / L of LVPA for 24,48,72 and 96 h, the growth inhibition rates of all the experimental groups were in a dose-dependent and time-dependent manner. The growth inhibition rate was 9% -66%, and the difference was statistically significant. P <0.001. The percentage of G1 phase in the control group was 57.8% -60.9%, while the experimental group showed G1 arrest at different degrees depending on the drug concentration and the duration of action. The G1 phase ratio was 58.2% 79.4%, compared with the control group, the difference was statistically significant, P <0.001. After VPA treatment, the expressions of Cyclin A, Cyclin D1 protein and mRNA were significantly down-regulated and the expression of p21Waf / cip1 protein and mRNA were significantly up-regulated in BGC-823 cells compared with the control group, P <0.001; There was no significant difference in mRNA expression between the two groups (P> 0.05). Conclusions: VPA, a specific histone deacetylase inhibitor, can significantly inhibit the proliferation of gastric cancer cells and arrest the cell cycle in G1 phase. It can inhibit the proliferation of gastric cancer cells by up-regulating the expression of p21Waf / cip1 mRNA and protein, down-regulating Cyclin D1, Cyclin A mRNA and protein expression are separately and / or synergistically achieved.