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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 effect of heterogeneous nuclear ribonucleoprotein A2B1 gene silencing on the apoptosis of 786-0 human renal cell carcinoma cell line and its possible mechanism, we constructed shRNA targeting hnRNPA2B1 gene, The recombinant plasmids were transfected into 786-0 cells. The silencing effects of recombinant plasmids on hnRNPA2B1 gene and the expression of Bcl-X was detected by RT-PCR. The changes of apoptosis were detected by flow cytometry. MTT colorimetric Method to detect cell proliferation. The results showed that compared with the control group, the gene silencing group apoptosis rate increased significantly, while the cell proliferation rate was significantly reduced (P <0.05). The level of Bcl-X (S) isoform mRNA in the silencing group was significantly increased, and the ratio of Bcl-X (S) / Bcl-X (L) was increased (P <0.05). hnRNPA2B1 gene silencing can promote the apoptosis of 786-0 renal carcinoma cells. The regulation of hnRNPA2B1 gene on the apoptosis is related to the expression of pro-apoptotic factor Bcl-X (S).