GRIM-19不同表达水平对前列腺癌细胞多西他赛化疗耐药性的影响

Effect of different expression levels of GRIM-19 on the resistance of prostate cancer cells to docetaxel chemotherapy

目的:探讨GRIM-19对多西他赛治疗前列腺癌(PCa)细胞的化疗耐药性的影响。方法:利用siRNA技术干扰PCa细胞的基因表达,建立GRIM-19过表达/敲除的PCa细胞模型,通过qPCR、Western印迹和流式细胞术等方法,探索GRIM-19不同表达水平对多西他赛诱导PCa细胞死亡的影响情况,揭示GRIM-19在PCa细胞化疗耐药性中的价值。结果:实验显示,GRIM-19在PCa组织和细胞中表达下调;GRIM-19敲除后,siGRIM-19在PC-3和LNCaP细胞中的表达明显降低,PC-3和LNCaP细胞接受不同剂量的多西他赛处理时,与对照组相比,GRIM-19的敲除降低了细胞死亡率;实验发现,GRIM-19过表达转染后,GRIM-19 mRNA和蛋白的表达明显增加,过表达的GRIM-19以剂量依赖性的方式促进多西他赛诱导的PC-3和LNCaP细胞死亡;用流式细胞术分析多西他赛诱导PC-3和PC-3-R细胞在不同时间和不同浓度的凋亡情况,结果显示PC-3-R细胞比PC-3细胞凋亡减少。结论:GRIM-19具有明显的促进细胞凋亡作用,是PCa抑癌基因;GRIM-19过表达促进了多西他赛诱导的PCa细胞死亡,提高了化疗敏感性。

Objective:To investigate the effect of GRIM-19 on the resistance of carcinoma cells to the chemotherapeutic agent docetaxel in the treatment of PCa.Methods:Using siRNA technology to interfere with the gene expression in PCa cells,we established a model of GRIM-19 overexpression/knockdown in PCa cells.We investigated the effect of different expression levels of GRIM-19 on docetaxel-induced death of the PCa cells by qPCR,Western blot and flow cytometry,and assessed the value of GRIM-19 in reducing the chemotherapy-resistance of PCa cells.Results:GRIM-19 was down-regulated in PCa tissues and cells.Knockout of GRIM-19 significantly decreased the expression of siGRIM19 in the PC-3 and LNCaP cells,and reduced their death rate when treated with docetaxel compared with the control group.The expressions of GRIM-19 mRNA and protein were remarkably upregulated after transfection with GRIM-19,and the overexpressed GRIM-19 promoted the death of the PC-3 and LNCaP cells treated with docetaxel in a dose-dependent manner.Flow cytometry analysis showed a lower apoptosis rate of PC-3-R cells than that of PC-3 cells at different time points of docetaxel-induction at different doses.Conclusion:GRIM-19 is a PCa suppressor gene with a significant facilitating effect on the apoptosis of PCa cells,and the overexpression of GRIM-19 promotes docetaxel-induced PCa cell death and improves the sensitivity of chemotherapy.