CAJ | 학술논문

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。
목적：탐토RNA간우침묵HDAC1기인대인류위암MGC803세포증식、조망급조단백을선화수평적영향。방법설계침대HDAC1기인소간우RNA편단，경지질체전염입MGC803세포，RT-PCR급Western blot감정기간우효과；MTT법회제세포생장곡선，TUNEL분석세포조망，Western blot검측조단백H3、H4을선화、조단백H3K9갑기화적영향급조망상관단백BCL-2、pro-caspase-9、pro-caspase-3、C-myc단백표체。결과 HDAC1 siRNA전염MGC803세포후，HDAC1기인적mRNA、단백표체균명현하강，유통계학의의（P<0.05）；60、120、240 nmol/L HDAC1 siRNA처리24 h후，세포조망솔분별위（18.5±3.5）%、（41.4±4.3）%、（59.2±5.5）%，이대조조부위（4.8±2.7）%，유통계학의의（P<0.05）；동시하조Bcl-2、proCaspase-9、proCaspase-3、c-Myc적표체；상조세포주기단백P21적표체；간우HDAC1기인가상조조단백H3，H4을선화수평，하조H3K9갑기화수평。결론간우HDAC1기인표체후가능통과상조여기인전록격활유관적조단백H3、H4을선화수평，하조여기인전록억제유관H3K9갑기화수평，종이억제세포증식，유도세포조망。
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.