RNA干扰Geminin基因表达对脑胶质瘤恶性生物学行为的影响  

Effects of RNA interference with Geminin gene expression on glioma biological behaviors

作　　者：王燕[1] 田薇[1] 章建国[1] 刘益飞[1] 靳钦[1] WANG Yan;TIAN Wei;ZHANG Jianguo;LIU Yifei;JIN Qin(Department of Pathology,Affiliated Hospital of Nantong University,Nantong 226001,China)

机构地区：[1]南通大学附属医院病理科,江苏南通226001

出　　处：《南京医科大学学报（自然科学版）》2021年第1期16-21,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南通市市级科技计划项目(MSZ18104)。

摘　　要：目的:探讨Geminin基因沉默对人脑胶质瘤细胞恶性生物学行为的影响及可能的发生机制。方法:选取南通大学附属医院脑外科2018—2019年手术切除的20例神经胶质瘤患者的新鲜标本,qRT-PCR结合基因表达谱数据动态分析(gene expression profilling interactive analysis,GEPIA)数据库分析胶质瘤组织中Geminin基因的表达;设计有效靶向沉默Geminin基因的siRNA序列和阴性对照序列,分别转染胶质瘤细胞U251,作为si-Geminin组和si-NC组,48 h后,qRT-PCR检测各组中Geminin mRNA的表达情况;分别采用CCK-8法、Transwell实验、Western blot法检测各组细胞的增殖情况、迁移和侵袭能力及Geminin、HBO1、Cdt1的蛋白表达水平。结果:qRT-PCR及GEPIA数据库分析结果显示脑胶质瘤组织中Geminin的表达明显高于瘤旁组织;siRNA干扰Geminin基因能显著抑制其在U251细胞中的表达;与control组及si-NC组相比,si-Geminin组细胞的增殖能力、细胞迁移和细胞侵袭的数量均显著降低,Geminin、HBO1及Cdt1蛋白表达水平显著降低。结论:Geminin在脑胶质瘤中高表达,敲除其表达可能通过下调HBO1及Cdt1抑制U251细胞的增殖、迁移和侵袭,Geminin有望成为人脑胶质瘤治疗新的分子靶点。Objective:To explore the effects of Geminin gene silencing on the biological behavior of human gliomas and the possible mechanism. Methods:Totally 20 pairs of fresh glioma specimens collected from patients who received a surgical resection in the Department of Brain Surgery in the Affiliated Hospital of Nantong University from 2018 to 2019 were analyzed by qRT-PCR combining GEPIA database to study the expression of Geminin gene in glioma tissues. The siRNA sequences that effectively targeted Geminin gene and the negative control sequences were designed and transfected into glioma U251 cells using LipofectamineTM 2000 reagent. The two groups were defined as siRNA-Geminin group and siRNA-NC group,respectively. In 48 h after transfection,qRT-PCR was used to detect the expressions of Geminin mRNA in each group. Then CCK8 method,Transwell assay,and Western blot were used to detect the cell proliferation,migration and invasion ability and the levels of Geminin,HBO1 and Cdt1 protein expression respectively. Results:Results of both qRT-PCR and GEPIA database analyses showed that the expression of Geminin mRNA in glioma tissues was significantly higher than that in adjacent tissue. The siRNA interference in Geminin gene could significantly inhibit its expression in U251 cells. Compared with siRNA-NC group,the cell proliferation ability,cell migrationand cell invasion quantity of the siRNA-Geminin group were significantly reduced,and the expression levels of Geminin,HBO1,and Cdt1 proteins were significantly lowered.Conclusion:Geminin is highly expressed in glioma. Knockdown of Geminin could inhibit the proliferation,migration,and invasion of U251 cells,possibly by down-regulating HBO1 and Cdt1. Thus,Geminin may be expected to be a new molecular target for the treatment of human glioma.

关 键 词：GEMININ 胶质瘤 SIRNA HBO1 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]