靶向抑制DNAJC1基因对多发性骨髓瘤细胞凋亡和增殖的影响

Influence of targeted inhibition of DNAJC1 gene on apoptosis and proliferation in multiple myeloma cells

目的研究DNAJC1基因对多发性骨髓瘤细胞增殖及凋亡的影响,初步探讨其作用机制。方法将DNAJC1-siRNA及阴性对照siRNA转染人多发性骨髓瘤细胞株U266,利用RT-qPCR及Western blot验证DNAJC1基因干扰效果,CCK-8检测DNAJC1基因对U266细胞增殖的影响,流式细胞仪检测其对细胞凋亡的影响。RT-qPCR及Western blot检测U266细胞中PTEN mRNA及蛋白表达。结果干扰组DNAJC1基因mRNA及蛋白表达明显降低,U266细胞的增殖明显受抑,抑制率为(58.95±8.13)%,与阴性siRNA组相比差异有统计学意义(P<0.05)。干扰组细胞凋亡率为(44.95±6.06)%,明显高于阴性siRNA组[(3.91±0.38)%]及对照组[(3.97±0.43)%,P<0.05]。与阴性siRNA组相比,干扰组PTEN的mRNA及蛋白表达明显升高(P<0.05)。结论靶向抑制DNAJC1基因可促进多发性骨髓瘤细胞的凋亡,抑制其增殖,这种作用可能是通过上调PTEN水平来实现的。

Objective To determine the effect of DNAJC1 gene on the proliferation and apoptosis of multiple myeloma cell lines and preliminarily investigate its mechanism. Methods DNAJC1-siRNA and its negative control(siRNA) were transfected into human multiple myeloma cell line U266 respectively. The interference efficiency was determined by RT-qPCR and Western blotting, and cell proliferation and apoptotic rate were measured with CCK-8 assay and flow cytometry, respectively. In addition, RT-qPCR and Western blotting were also used to examine the mRNA and protein expression of PTEN in U266 cells. Results The mRNA and protein levels of DNAJC1 were downregulated in the interference group. The proliferation of U266 cells was inhibited significantly in the interference group, with an inhibitory rate of(58.95±8.13)%, apparently higher than that of the negative siRNA group(P<0.05). The apoptotic rate was(44.95±6.06)% in the interference group, which notably exceeded that of the negative siRNA group [(3.91±0.38)%] and the control group [(3.97±0.43)%](P<0.05). The mRNA and protein levels of PTEN were significantly increased in the interference group(P<0.05). Conclusion Targeted inhibition of DNAJC1 gene can promote the apoptosis and inhibit the proliferation in multiple myeloma cells, which may be mediated by up-regulation of PTEN.