南方医科大学学报

南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (02): 246-.

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RNA干扰沉默HDAC1基因对胃癌细胞增殖、凋亡、组蛋白乙酰化和甲基化的影响

庄涵虚，马旭东，赖亚栋，许向农，王小众   

  • 出版日期:2014-02-20 发布日期:2014-02-20

Effect of siRNA targeting HDAC1 gene on proliferation, apoptosis, histone acetylation,
and histone metylation in gastric cancer cells in vitro

  • Online:2014-02-20 Published:2014-02-20

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摘要： 目的探讨RNA干扰沉默HDAC1基因对人类胃癌MGC803细胞增殖、凋亡及组蛋白乙酰化水平的影响。方法设计针对
HDAC1基因小干扰RNA片段，经脂质体转染入MGC803细胞，RT-PCR及Western blot鉴定其干扰效果；MTT法绘制细胞生长
曲线，TUNEL分析细胞调亡，Western blot 检测组蛋白H3、H4 乙酰化、组蛋白H3K9 甲基化的影响及凋亡相关蛋白BCL-2、
pro-caspase-9、pro-caspase-3、C-myc蛋白表达。结果HDAC1 siRNA转染MGC803细胞后，HDAC1基因的mRNA、蛋白表达均
明显下降，有统计学意义（P<0.05）；60、120、240 nmol/L HDAC1 siRNA处理24 h 后，细胞凋亡率分别为（18.5±3.5）%、（41.4±
4.3）%、（59.2±5.5）%，而对照组仅为（4.8±2.7）%，有统计学意义（P<0.05）；同时下调Bcl-2、proCaspase-9、proCaspase-3、c-Myc的
表达；上调细胞周期蛋白P21的表达；干扰HDAC1基因可上调组蛋白H3，H4乙酰化水平，下调H3K9甲基化水平。结论干扰
HDAC1基因表达后可能通过上调与基因转录激活有关的组蛋白H3、H4乙酰化水平，下调与基因转录抑制有关H3K9甲基化水
平，从而抑制细胞增殖，诱导细胞凋亡。

Abstract: Objective To investigate the effect of silencing histone deacetylases1 (HDAC1) gene by RNA interference on the
proliferation, apoptosis and histone modulation in gastric cancer MGC-803 cell line. Methods The optimal segment targeting
HDAC1 gene was designed and transfected into MGC-803 cells by Lipofectamine TM2000. HDAC1 mRNA and protein in the
transfected cells were detected by RT-PCR and Western blotting, respectively. The growth inhibition of MGC803 cells was
evaluated by MTT assay and the cell apoptosis was detected with TUNEL assay. The expression of Bcl-2, procaspase-9,
procaspase-3, c-Myc, histone acetylation of H3, H4, and histone methylation of H3K9 was detected by Western blotting.
Results The siRNA targeting HDAC1HDAC1 markedly suppressed mRNA expression, inhibited cell proliferation and induced
apoptosis of MGC-803 cells in a concentration manner. Transfection of the cells with HDAC1 siRNA at 0, 30, 60, and 120 nmol/L
for 24 h resulted in a cell apoptotic rate of (4.8 ±2.7)%, (18.5 ±3.5)%, (41.4 ±4.3)%, and (59.2 ±5.5)%, respectively, and caused
down-regulation of the expressions of Bcl-2, proCaspase9, proCaspase3 and c-Myc, upregulation of histone acetylation of H3,
H4, and down-regulation of histone methylation of H3K9. Conclusion Silencing HDAC1 gene expression with HDAC1 siRNA
can promote histone H3 and H4 acetylation and inhibit histone methylation of H3K9 to suppress the proliferation and induce
apoptosis of gastric cancer MGC-803 cells.