USP7抑制剂促进耐替莫唑胺的原代胶质母细胞瘤细胞凋亡的研究

Application of USP7 inhibitors to promote apoptosis in temozolomide-resistant primary glioblastoma cells

目的研究USP7的抑制剂P5091对耐TMZ的原代胶质母细胞瘤的影响.方法用免疫组化法分析不同初发胶质母细胞瘤组织和复发胶质母细胞瘤中USP7的表达情况.用Western Blot检测初发胶质母细胞瘤组织和复发胶质母细胞瘤组织中USP7的表达差异.用Western Blot检测原代胶质母细胞瘤细胞SHG-140和其TMZ耐药株SHG-140R中USP7的表达差异.用CCK8检测P5091对SHG-140和SHG-140R存活率的影响.用流式细胞术检测P5091对SHG-140和SHG-140R细胞凋亡的影响.用Western Blot检测P5091治疗后SHG-140R中细胞凋亡蛋白的表达.结果胶质母细胞瘤中的USP7表达高于正常脑组织(P<0.01),不同初发胶质母细胞瘤组织和复发胶质母细胞瘤中,USP7的表达差异不明显.相比SHG-140,应用P5091后SHG-140R存活率下降更明显,流式细胞术结果提示应用P5091后SHG-140R细胞凋亡更显著(P<0.01).Western Blot结果显示应用P5091后SHG-140R细胞中细胞凋亡蛋白CLEAVED-PARP、CLEAVED-CASPASE9、CLEAVED-CASPASE3表达均明显升高(P<0.05,P<0.01).结论P5091能够促进耐TMZ的原代胶质母细胞瘤细胞凋亡.

Objective To study the effect of applying P5091,an inhibitor of USP7,on TMZ-resistant primary glioblastoma.Methods The expression of USP7 in different primary glioblastoma tissues and recurrent glioblastoma was analyzed by immunohistochemistry.Differences in USP7 expression in primary glioblastoma tissues and recurrent glioblastoma tissues were detected by Western Blot.Differences in USP7 expression in primary glioblastoma cells SHG-140 and its TMZ-resistant strain SHG-140R were detected by Western Blot.The effect of P5091 on the survival of SHG-140 and SHG-140R was detected by CCK8.The effect of P5091 on SHG-140 and SHG-140R apoptosis was detected by flow cytometry.The expression of apoptotic proteins in SHG-140R after P5091 treatment was detected by Western Blot.Results The expression of USP7 in glioblastoma was higher than that in normal brain tissues(P<0.01),and the difference of USP7 expression was not obvious in different primary glioblastoma tissues and recurrent glioblastoma.Compared with SHG-140,the survival rate of SHG-140R decreased more significantly after the application of P5091,and the flow cytometry results suggested that the apoptosis of SHG-140R cells was more significant after the application of P5091(P<0.01).Western Blot results showed that the apoptotic proteins CLEAVED-PARP,CLEAVED-CASPASE9,and CLEAVED-CASPASE3 expression were significantly elevated in SHG-140R cells(P<0.05,P<0.01).Conclusion P5091 can promote apoptosis in TMZ-resistant primary glioblastoma cells.