HDAC2 siRNA转染对人胰腺癌细胞增殖及凋亡的影响

摘要

目的:探讨RNA干扰沉默组蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)基因表达对人胰腺癌细胞增殖和凋亡的影响.方法:培养人胰腺癌PaTu8988细胞株,并将其随机分为5组:空白对照组,20 nmol/L HDAC2 siRNA阴性组,40 nmoL/L HDAC2 siRNA阴性组,20 nmol/L HDAC2 siRNA组,40 nmol/L HDAC2 siRNA组.合成针对HDAC2基因的小干扰RNA (small interfering RNA,siRNA),利用阳离子脂质体将HDAC2 siRNA瞬时转染人胰腺癌PaTu8988细胞,分别采用实时荧光定量PCR (qRT-PCR)和蛋白质印迹法检测HDA C2基因及蛋白的表达;MTT比色法检测细胞的增殖率;流式细胞术测定细胞凋亡率.结果:与空白对照组和阴性siRNA组对比,HDAC2 siRNA组的HDAC2基因和蛋白的表达水平均明显下降(P均＜0.05),细胞增殖率减慢(P＜0.01),凋亡率明显升高(P＜0.01).结论:采用RNA干扰沉默人胰腺癌细胞HDAC2基因表达,可抑制癌细胞增殖和诱导其凋亡.%To investigate the effects of siRNA-mediated histone deacetylase 2 (HDAC2) gene silence on the proliferation and apoptosis of human pancreatic cell line PaTu8988.Methods:Human pancreatic cancer cell line PaTu8988 cells were cultured and divided into five groups:control group,20 nmol/L negative siRNA group,40 nmol/L negative siRNA group,20 nmol/L HDAC2 siRNA group,and 40 nmoL/L HDAC2 siRNA group.Transient transfection of small interfering RNA (siRNA) against HDAC2 on human pancreatic cancer cell line PaTu8988 cells were performed by Lipofectamine 2000.Expression of HDAC2 gene was detected by quantitative real-time PCR and western blotting,respectively.The cell proliferation was determined by MTT assay.Flow cytometry was used to examine the cells apoptosis.Results:Compared with control group and negative siRNA groups,HDAC2 siRNA groups had markedly lower levels of HDAC2 expressions and lower cell growth rate,but higher cell apoptosis rate(P ＜0.05 or P ＜0.01).Conclusion:Transfection of HDAC2 siRNA could effectively inhibit the expression of HDAC2 in PaTu8988 cell lines,which suppressed the proliferation and induced apotosis of PaTu8988 cells.