IDH1-R132H突变体通过调节HIF-1α通路抑制胶质瘤U87细胞恶性进展  

The IDH1-R132H mutant inhibits malignant progression of glioma U87 cells by regulating the HIF-1α pathway

作　　者：方松 唐国强[1] 叶友忠[1] 李明松 陈加贝 FANG Song;TANG Guoqiang;YE Youzhong;LI Mingsong;CHEN Jiabei(Department of Neurosurgery,Chenzhou First People's Hospital,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院神经外科,郴州423000

出　　处：《临床神经外科杂志》2024年第6期654-659,共6页Journal of Clinical Neurosurgery

基　　金：2020年湘南学院校级科学研究项目(2020XJ133);2024年湘南学院校级科学研究项目(2024XJ124)。

摘　　要：目的 探究异柠檬酸脱氢酶1(IDH1)-R132H突变体通过调节低氧诱导因子-1α(HIF-1α)通路抑制胶质瘤U87细胞恶性进展。方法 实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白免疫印迹法检测胶质瘤组织和细胞IDH1 mRNA和蛋白表达水平。将U87细胞分为对照组、Vector组、IDH1 wt组、IDH1-R132H组,qRT-PCR和Western Blot检测IDH1 wt、IDH1-R132H转染效率,噻唑蓝(MTT)和克隆形成实验检测细胞增殖,Transwell检测细胞迁移,流式细胞仪检测细胞凋亡,Western Blot检测IDH1、PCNA、基质金属蛋白酶-2(MMP-2)、Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)、血管内皮生长因子(VEGF)、HIF-1α、HIF-2α蛋白表达。结果 与癌旁组织和正常胶质细胞NHAs相比,胶质瘤组织和人神经胶质母细胞瘤细胞T98G、U138、U87中IDH1 mRNA和蛋白表达水平明显降低(P<0.05)。qRT-PCR和Western Blot结果表明,IDH1 wt、IDH1-R132H转染成功;与对照组或Vector组比较,IDH1-R132H组细胞活性、PCNA、MMP-2、Bcl-2蛋白表达明显降低,VEGF、HIF-1α、HIF-2α蛋白表达明显升高,且IDH1-R132H组克隆细胞数和迁移细胞数减少,细胞凋亡率、Bax蛋白表达明显增加(P<0.05);但对照组、Vector组、IDH1 wt组上述指标两两比较,差异均无统计学意义(P>0.05)。结论 IDH1-R132H突变体可能通过调节HIF-1α通路抑制胶质瘤U87细胞增殖和迁移,并诱导细胞凋亡。Objective To investigate the inhibition of malignant progression of glioma U87 cells by the regulation of hypoxic-induced-factor-1α(HIF-1α)pathway by mutant isocitrate dehydrogenase 1 R132H(IDH1-R132H).Methods Real-time quantitative fluorescent polymerase chain reaction(qRT-PCR)and Western Blot were used to detect IDH1 mRNA and protein expression levels in glioma tissues and cells.U87 cells were divided into control group,Vector group,IDH1-wt group and IDH1-R132H group.The transfection efficiency of IDH1-wt and IDH1-R132H were detected by qRT-PCR and Western Blot,and cell proliferation was detected by MTT and clonal formation experiments.Cell migration was detected by Transwell,apoptosis was detected by flow cytometry,Western Blot analysis of IDH1,PCNA,matrix metalloproteinase-2(MMP-2),Bcl-2 associated X protein(Bax),B-cell lymphoma/leukemia-2(Bcl-2),vascular endothelial growth factor(VEGF),HIF-1α,HIF-2αprotein expression.Results The expression levels of IDH1 mRNA and protein in glioma tissues and human glioblastoma cells T98G,U138,U87 were significantly lower than those in para-cancerous tissues and normal glioblastoma cells(P<0.05).qRT-PCR and Western Blot proved successful transfection of IDH1-wt and IDH1-R132H.Compared with control group or Vector group,IDH1-R132H group significantly decreased cell activity,PCNA,MMP-2 and Bcl-2 protein expression,significantly increased VEGF,HIF-1αand HIF-2αprotein expression,decreased the number of cloned cells and migrated cells,and significantly increased apoptosis rate and Bax protein expression,and the number of cloned cells and migrated cells in IDH1-R132H group were decreased,while the apoptosis rate and Bax protein expression were significantly increased(P<0.05).However,pairwise comparison of the above indicators in control group,Vector group and IDH1-wt group showed no statistical significance(P>0.05).Conclusions The IDH1-R132H mutant may inhibit the proliferation and migration of glioma U87 cells and induce apoptosis by regulating the HIF-1αpathway.

关 键 词：IDH1-R132H突变体 HIF-1α通路 胶质瘤 细胞增殖 细胞迁移 细胞凋亡 

分 类 号：R739.41[医药卫生—肿瘤]