β-TrCP1和HAUSP影响胶质母细胞瘤细胞增殖和侵袭并调控UHRF1蛋白水平  

UHRF1β-TrCP1 and HAUSP affect the proliferation and invasion abilities of glioblastoma and regulate UHRF1 protein level

作　　者：李玉辉[1] 赵喜庆[1] 陆丽娟 刘艳坤[3] 刘岩 胡万宁[3] 李玉凤[3] Li Yuhui;Zhao Xiqing;Lu Lijuan;Liu Yankun;Liu Yan;Hu Wanning;Li Yufeng(Department of Neurosurgery,Tangshan People’s Hospital,Tangshan 063001,China;Graduate School of North China University of Science and Technology,Tangshan 063210,China;Cancer Institute,Tangshan People’s Hospital,Tangshan 063001,China;Department of Bioengineering,College of Life Sciences,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]唐山市人民医院神经外科,063001 [2]华北理工大学研究生院,唐山063210 [3]唐山市人民医院肿瘤研究所,063001 [4]华北理工大学生命科学院生物工程系,唐山063210

出　　处：《中华神经创伤外科电子杂志》2019年第4期233-238,共6页Chinese Journal Of Neurotraumatic Surgery：Electronic Edition

基　　金：唐山市科技创新团队培养计划项目（17130205D)

摘　　要：目的分析β-转导重复相容蛋白(β-TrCP1)和疱疹病毒相关性泛素特异性蛋白酶(HAUSP)对胶质母细胞瘤增殖和侵袭能力以及表观遗传调控因子UHRF1表达的影响。方法胶质母细胞瘤U87细胞内分别转染N1空质粒(NC组)和β-TrCP1-N1过表达质粒(β-TrCP1组),以及分别转染无意义siRNA序列(siRNA阴性对照,si-NC组)和靶向HAUSP mRNA的siRNA(si-HAUSP组)。RT-qPCR法和Western blot法分别检测各组细胞中β-TRCP1、HAUSP和UHRF1的mRNA以及蛋白水平。CCK8法和基质胶-transwell侵袭实验分别检测各组细胞的增殖和侵袭能力。结果质粒转染后第4天和第5天,β-TrCP1组细胞的增殖能力显著低于同时期的NC组细胞(均P<0.05);转染第4天,β-TrCP1组穿过基质胶的细胞数显著低于NC组细胞(P<0.05)。转染siRNA后第4天和第5天,si-HAUSP组细胞的增殖能力显著低于同时期si-NC组细胞(P<0.05);转染si-HAUSP4d,si-HAUSP组穿过基质胶的细胞数显著低于si-NC组(P<0.05)。此外,过表达β-TrCP1和敲低HAUSP均不影响UHRF1的mRNA水平(均P>0.05),但均可降低UHRF1蛋白水平(均P<0.05)。结论过表达β-TrCP1和敲低HAUSP均可抑制胶质母细胞瘤U87细胞的增殖和侵袭能力,同时降低UHRF1蛋白水平。UHRF1蛋白水平改变可能是β-TrCP1和HAUSP影响胶质母细胞瘤增殖和侵袭能力的机制之一。Objective To analyze the effects of beta-transducin repeat-containing proteins (β-TrCP1) and herpes associated ubiquitin specific proteas (HAUSP) on proliferation and invasion abilities of glioblastoma cells, as well as the expression of epigenetic factor UHRF1. Methods The U87 glioblastoma cells were respectively transfected with blank N1 plasmid (NC) and N1 plasmid containing β-TrCP1 coding sequence (β-TrCP1), as well as control oligonucleotides (si-NC) and siRNAs targeting HAUSP mRNA (si-HAUSP). The mRNA and protein levels of β-TrCP1, HAUSP and UHRF1 were detected by RT-qPCR and Western blot, respectively. The proliferation and invasion abilities of each group cells were detected by CCK8 and Matrigel-transwell invasive assay, respectively. Results Post-transfection of plasmids, the proliferation abilities of β-TrCP1 group cells were markedly decreased compared with those of NC groups cells at the fourth and the fifth day post-transfection (all P<0.05). The number of invased cells of β-TrCP1 group was significantly lower than that of NC group at the fourth day post-transfection (P<0.05). On the other hand, post-transfection of siRNAs, the proliferation abilities of si-HAUSP group cells were significantly decreased compared with those of si-NC groups cells at the fourth and the fifth day post-transfection (P<0.05). The number of invasive cells of si-HAUSP group was significantly lower than that of si-NC group at the fourth day post-transfection (P<0.05). Furthermore, the protein level of UHRF1 was markedly downregulated by β-TrCP1 overexpression or HAUSP knockdown (all P<0.05), but the mRNA level was not affected (all P>0.05). Conclusion Both the proliferation and invasion abilities of glioblastoma U87 cells, as well the UHRF1 protein level, could be inhibited by β-TrCP1 overexpression or HAUSP knockdown. The affection of β-TrCP1 and HAUSP on proliferation and invasion abilities of GBM is probably mediated by the protein level change of UHRF1.

关 键 词：胶质母细胞瘤 β-转导重复相容蛋白 疱疹病毒相关性泛素特异性蛋白酶 UHRF1 

分 类 号：R739.4[医药卫生—肿瘤]