沉默hnRNPA2B1基因促进肾癌细胞凋亡

Gene Silencing on hnRNPA2B1 Promotes Apoptosis of Renal Cell Carcinoma Cells

为观察不均一型核糖核蛋白A2B1(heterogeneous nuclear ribonucleoprotein A2B1,hnRNPA2B1)基因沉默对786-0肾癌细胞株凋亡的影响及其可能的机制,构建了针对hnRNPA2B1基因的短发夹RNA(shRNA)重组质粒,并转染786-0细胞;RT-PCR检测重组质粒对hnRNPA2B1基因的沉默效果以及对凋亡因子Bcl-X亚型表达的影响;流式细胞计数检测细胞凋亡改变;MTT比色法检测细胞增殖能力。结果显示,与对照组相比,基因沉默组细胞凋亡率明显增加,而细胞增殖率显著降低(P<0.05)。沉默组中促凋亡因子Bcl-X(S)亚型mRNA明显增加,Bcl-X(S)/Bcl-X(L)比值增大(P<0.05)。hnRNPA2B1基因沉默促进786-0肾癌细胞凋亡,hnRNPA2B1基因对凋亡的调节与影响促凋亡因子Bcl-X(S)的表达有关。

To investigate the effects of hnRNPA2BI gene silencing on apoptosis of 786-0 cells and its potential mechanism, hnRNPA2B1 shRNA recombinant plasmids were constructed and transfected into renal cell carcinoma 786-0 cells. The effects of hnRNPA2B1 shRNA recombinant plasmids on hnRNPA2B1 gene silencing and the expression of BcI-X isoforms were detected by RT-PCR. The apoptosis of 786-0 cells were assayed by flow cytometry. MTT method was used to observe the proliferation ability of 786-0 cells. Compared to the control groups, the apoptosis rates of gene-silencing groups were increased significantly while the cell multiplication rates were notably decreased （P〈0.05）; The mRNA expression levels of pro-apoptotic factor Bcl-X（S） and the ratio of BcI-X（S）/Bcl-X（L） were increased significantly after the hnRNPA2B1 gene have been silenced （P〈0.05）. In conclusion, hnRNPA2B1 gene silencing could effectively promote apoptosis of 786-0 cells, the mechanism of which probably relates to its affection on the expression of Bcl-X（S）.