ING1基因沉默后通过抑制Caspase 2调节胃腺癌AGS细胞凋亡

INGl Plays an Important Role in Regulating AGS Cell Apoptosis by Down-regulated Caspase 2 Gene Expression

目的:探讨RNA干扰沉默ING1基因表达对胃腺癌细胞AGS细胞凋亡的影响机制。方法:将体外合成的ING1基因特异性的siRNA利用脂质体2 000转染到胃腺癌细胞AGS,应用流式细胞技术和TUNEL技术检测ING1基因表达沉默对AGS细胞凋亡的影响应用荧光定量PCR技术和Western blot技术检测转染后40 hAGS细胞中ING1、Caspase 2和Caspase 3的表达。结果:转染后40 h,体外合成的siRNA对AGS细胞的ING1表达抑制作用明显。转染前凋亡率为(11.06±0.97)%,转染40 h后凋亡率为(6.70±0.41)%,转染前后凋亡率的改变差异有统计学意义(P<0.05)。转染后40 h AGS细胞中ING1、Caspase 2和Caspase 3的mRNA相对表达分别为0.170 7±0.06,0.125±0.03和0.999±0.10。转染后40 hAGS细胞中ING1、Caspase 2和Caspase 3的蛋白质水平改变的趋势与mRNA水平一致。结论:RNA干扰沉默ING1基因表达后,AGS细胞中Caspase 2表达明显下调,ING1基因沉默后通过抑制Caspase 2调节胃腺癌细胞AGS细胞凋亡。

To assess the mechanism of ING1 gene silencing on cell apoptosis of gastric cancer cell line AGS. Methods： The synthetic siRNA specific to ING1 was transfected into AGS cells. Cell apoptosis was evaluated by flow cytometry and TUNEL. The expression levels of ING1, caspase 2 and caspase 3 were detected by real-time PCR and western-blot. Results： The synthetic siRNA could effectively inhibit the expression of ING1 in AGS cells. The apoptosis cells number of ING1 knocked-down group was 6.70 ± 0.41%,which was significantly decreased compared with the control group （ 11.06 ± 0.97%, P 〈 0. 05 ）. 40 hours after transfection, the mRNA expression of ING1, caspase 2 and caspase 3 was 0.1707 ± 0.06, 0.125 ± 0.03 and 0.999 ± 0.10, respectively. The tendency of protein variation of ING1, caspase2 and caspase 3 was consistent with mRNA. Conclusion： ING1 gene silencing could down-regulated caspase 2 gene expression and ING1 plays an important role in regulating AGS cell apoptosis by down-regulated caspase 2 gene expression.