摘要
目的长链非编码RNA(lncRNA) MTHFD2基因在胶质母细胞瘤(GBM)组织与正常脑组织中的表达有明显差异,但其在肿瘤尤其是GBM的生物学功能目前尚不清楚。文中旨在研究lncRNA MTHFD2在人GBM组织和细胞系中的表达情况,并观察下调其表达对GBM细胞生物学功能的影响。方法选取2017年9月至2017年12月东部战区总医院神经外科经手术切除的9例GBM患者标本,同时取其配对的瘤旁组织作为正常对照组织。取对数生长期的U251和U-87MG细胞,接种细胞24 h后分别加入LV-MTHFD2-shRNA病毒液(U251shRNA组、U-87MGshRNA组)和空载LV-control病毒液(U251对照组、U-87MG对照组)。采用qRT-PCR检测GBM组织和细胞系中lncRNA MTHFD2的表达。CCK-8检测细胞增殖和对化疗药物替莫唑铵的耐药情况。Transwell小室评价细胞迁移能力的变化等。结果 GBM组织lncRNA MTHFD2相对表达量明显高于正常对照组织[(5.13±3.96)vs(1.27±0.58)],差异有统计学差异(P<0.05)。与U251对照组MTHFD2的相对表达量(1.02±0.08)比较,U251shRNA组(0.05±0.01)明显降低(P<0.01);U-87MGshRNA组较U-87MG对照组亦明显降低(P<0.05)。U251shRNA组穿过Transwell微孔滤膜的细胞数量与U251对照组比较明显降低[(41.4±6.99)个/视野vs (125.8±25.27)个/视野],差异有统计学意义(P<0.01);U-87MGshRNA组较U-87MG对照组亦明显降低(P<0.05)。CCK-8结果显示,在第4天,U251shRNA组A值较U251对照组明显降低,U-87MGshRNA组A值较U-87MG对照组亦明显降低(P<0.05)。细胞耐药性结果显示:U251shRNA组细胞抑制率明显高于U251对照组,U87-MGhRNA组亦明显低于U87-MG对照组(P<0.05)。结论 lncRNA MTHFD2在GBM细胞中的表达下调可降低细胞的增殖和迁移能力,增强对化疗药物替莫唑铵的敏感性,提示其可能为GBM潜在的治疗靶标。
Objective The long non-coding RNA(lncRNA)MTHFD2 gene is expressed differentially in glioblastoma(GBM)and normal brain tissues,but its biological role in tumors,and particularly in GBM,remains unclear.This study aims to investigate the expression of lncRNA MTHFD2 in the GBM tissue and four GBM cell lines,and explore the effect of its down-regulated expression on the biological function of GBM cells.Methods Specimens of GBM and the paracancerous tissue(as normal control)were collected from 9 patients treated by surgical resection in our Department of Neurosurgery between September and December 2017 LV-MTHFD2-shRNA(U251 shRNA and U-87MG shRNA groups)and empty LV-control solution(U251 shRNA and U-87MG control groups)were transfected into the U251 and U-87MG cell lines.The expressions of lncRNA MTHFD2 in the GBM tissue and the GBM cell lines were detected by qRT-PCR,the chemosensitivity and proliferation of the cells after transfection measured by CCK-8 assay,and the changes in the cell migration ability determined by Transwell assay.Results The relative expression of lncRNA MTHFD2 was significantly higher in the GBM than in the normal tissue(5.13±3.96 vs 1.27±0.58,P<0.05),while that of MTHFD2 was remarkably lower in the U251 shRNA than in the U251 control group(0.05±0.01 vs 1.00±0.00,P<0.01),and so was that in the U-87MG shRNA than in the U-87MG control(P<0.05).The number of cells penetrating the Transwell membrane was markedly lower in the U251 shRNA group than in the U251 control(41.4±6.99 vs 125.8±25.27 per field of view,P<0.01),and so was that in the U-87MG shRNA than in the U-87MG control(P<0.05).CCK-8 assay showed that,at 4 days after transfection,the A value was significantly decreased in the U251 shRNA and U-87MG shRNA groups as compared with the U251 control and U-87MG control groups(P<0.05).Cellular drug resistance test manifested remarkably reduced fifty percent inhibitory concentrations(IC50)in the U251 shRNA and U-87MG shRNA groups as compared with the U251 control and U-87MG control groups(P<0.05).Conclusion DDown-regulation of the expression of lncRNA MTHFD2 can inhibit the proliferation and migration of U251 and U-87MG cells and enhance the chemosensitivity of the cells to temozolomide,which suggests that lncRNA MTHFD2 could be a potential therapeutic target against GBM.
作者
韩艳玲
周梦良
王汉东
HAN Yan-ling;ZHOU Meng-liang;WANG Han-dong(Department of Neurosurgery,General Hospital of Eastern Theater Command,PLA,Nanjing 210002,Jiangsu,China)
出处
《医学研究生学报》
CAS
北大核心
2019年第4期369-373,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金(81672503)
作者简介
韩艳玲,理学硕士;通信作者:王汉东,E-mail:njhdwang@hotmail.com.
