细胞周期相关基因的胶质母细胞瘤预后模型构建及RFC2的细胞增殖效应研究  

Establishment of a prognostic model for glioblastoma associated with cell cycle genes and study on the cell proliferation effect of RFC2

作　　者：王二静 吴伟 周浩宇 王乙錩 项健阳 王佳[1] 王茂德[1] WANG Erjing;WU Wei;ZHOU Haoyu;WANG Yichang;XIANG Jianyang;WANG Jia;WANG Maode(Department of Neurosurgery,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院神经外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2024年第5期748-756,共9页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省自然科学基金青年项目(No.2022JQ-810);陕西省自然科学基金面上项目(No.2021JM-264)。

摘　　要：目的研究复制因子C亚基2(RFC2)与胶质母细胞瘤(GBM)患者预后和肿瘤细胞增殖之间的关系,并探索其在GBM发展中的潜在分子通路。方法利用生物信息学方法,筛选出可作为GBM独立预后因素的细胞周期基因,结合临床指标,构建GBM患者风险评分模型并验证其预测能力,对目标基因RFC2进行GO、KEGG和GSEA分析。取对数生长期的U87 GBM细胞进行慢病毒转染后分组(空白对照组、shRFC2#1、shRFC2#2),通过qRT-PCR、Western blotting、Edu染色、克隆形成实验检测mRNA表达、蛋白表达和细胞增殖。结果RFC2在GBM中表达上调,并随着胶质瘤病理级别升高,呈明显上升趋势。基因功能与通路分析结果提示,RFC2在姐妹染色体分离、染色体分离、细胞器裂变、核分裂、核有丝分裂等进程中,促进细胞周期过程中G1向S期转变。qRT-PCR和Western blotting结果显示,相较空白对照组,慢病毒敲低组中转录mRNA减少(P<0.0001)、翻译蛋白量减少;同样,与空白对照组相比,慢病毒敲低组Edu染色阳性率降低(P<0.0001),集落形成能力相应降低(P<0.001)。结论RFC2在GBM中高表达,且与胶质瘤分级和患者不良预后相关,促进GBM细胞增殖;RFC2有可能成为GBM潜在的生物标志物和治疗靶标。Objective To investigate the relationship of replication factor C subunit 2(RFC2)with the prognosis of glioblastoma(GBM)and cell proliferation,as well as its underlying molecular pathway in GBM development.Methods Using bioinformatics methods,cell cycle genes were screened as independent prognostic factors for GBM.Combined with clinical indicators,a risk scoring model for GBM patients was established and validated.The target gene RFC2 was analyzed with GO,KEGG,and GSEA.U87 GBM cells at logarithmic growth stage were transfected with lentivirus and divided into different groups(control,ShRFC2#1,and shRFC2#2 groups).qRT-PCR,Western blotting,Edu staining,and cloning assay were used to detect mRNA expression,protein expression,and cell proliferation.Results The expression of RFC2 was upregulated in GBM and showed an obvious upregulation trend with the increase of pathological grade of glioma.The analyses of gene function and pathway indicated that RFC2 was involved in the processes of sister chromosome segregation,chromosome segregation,organelle fission,and mitosis by promoting the transition of G1 to S phase during cell cycle.qRT-PCR and Western blotting showed that compared with the control group,the amount of mRNA and translated protein in the knockdowned groups decreased(P<0.0001).The positive rate of Edu staining and the colony forming ability decreased(P<0.0001,P<0.001).Conclusion RFC2 is highly expressed in glioblastoma and associated with pathological grade of glioma and poor prognosis of patients.It also promotes the cell proliferation function of glioblastoma.RFC2 may be a potential biomarker and therapeutic target for glioblastoma.

关 键 词：胶质母细胞瘤(GBM) 复制因子C亚基2(RFC2) 预后 增殖 

分 类 号：R739.41[医药卫生—肿瘤]