Par-4基因过表达调节肾母细胞瘤顺铂敏感性的实验研究

Experimental study about the regulating effect of Par-4 gene overexpression on the nephroblastoma sensitivity to cisplatin

目的:研究Par-4基因过表达对肾母细胞瘤顺铂敏感性的调节作用。方法:培养肾母细胞瘤SKNEP-1细胞并分为四组,对照组用不含血清及药物的RMPI-1640处理,顺铂组用含有5μg/mL顺铂的无血清RMPI-1640处理,Par-4组用无血清RMPI-1640转染Par-4过表达质粒,顺铂+Par-4组用含有5μg/mL顺铂的无血清RMPI-1640转染Par-4过表达质粒。处理后测定细胞的增殖活力及促凋亡基因、迁移及侵袭基因的表达量。结果:不同条件处理后8h、16h、24h,顺铂组、Par-4组、顺铂+Par-4组的细胞增殖活力值均显著低于对照组,顺铂+Par-4组的细胞增殖活力值显著低于顺铂组、Par-4组;不同条件处理后24h,顺铂组、Par-4组、顺铂+Par-4组细胞中Bim、PDCD4、WT1、RGS4、Axin、KAI1、E-cadherin、PPARγ、PTEN的mRNA表达量均显著高于对照组,GDNF、GFRα1、TUBB3、NME1、FGF1的mRNA表达量均显著低于对照组;顺铂+Par-4组细胞中Bim、PDCD4、WT1、RGS4、Axin、KAI1、E-cadherin、PPARγ、PTEN的mRNA表达量显著高于顺铂组、Par-4组,GDNF、GFRα1、TUBB3、NME1、FGF1的mRNA表达量显著低于顺铂组、Par-4组。结论:Par-4基因过表达能够增加肾母细胞瘤对顺铂的敏感性,降低细胞增殖活力并促进细胞凋亡、抑制细胞迁移和侵袭。

Objective:To study the regulating effect of Par-4 gene overexpression on the nephroblastoma sensitivity to cisplatin.Methods:Nephroblastoma SK-NEP-1 cells were cultured and divided into four groups,control group were treated with RMPI-1640 without serum or drugs,cisplatin group were treated with serum-free RMPI-1640 containing 5μg/mL cisplatin,Par-4 group were transfected by Par-4 overexpression plasmids with serum-free RMPI-1640,and cisplatin+ Par-4 group were transfected by Par-4 overexpression plasmid with serum-free RMPI-1640 containing 5μg/mL cisplatin.The cell proliferation activity as well the expression of apoptosis genes,migration genes and invasion genes was measured.Results:Eight hours,16 hand 24 hafter different conditions of treatment,the cell proliferation activity of cisplatin group,Par-4 group and cisplatin+ Par-4 group were significantly lower than that of control group,and the cell proliferation activity of cisplatin+ Par-4 group was significantly lower than that of cisplatin group and Par-4 group;24 hafter different conditions of treatment,Bim,PDCD4,WT1,RGS4,Axin,KAI1,E-cadherin,PPARγand PTEN mRNA expression in cisplatin group,Par-4 group and cisplatin+Par-4 group were greatly higher than those in control group whereas GDNF,GFRα1,TUBB3,NME1 and FGF1 mRNA expression were greatly lower than those in control group;Bim,PDCD4,WT1,RGS4,Axin,KAI1,E-cadherin,PPARγand PTEN mRNA expression in cisplatin+ Par-4 group were greatly higher than those in cisplatin group and Par-4 group whereas GDNF,GFRα1,TUBB3,NME1 and FGF1 mRNA expression were greatly significantly lower than those in cisplatin group and Par-4 group.Conclusion:Par-4 gene overexpression can increase the nephroblastoma sensitivity to cisplatin,reduce cell proliferation activity,promote apoptosis and inhibit cell migration and invasion.