摘要
目的探讨miR-103a-3p过表达对胶质瘤C6细胞恶性生物学行为的影响及对裸鼠移植瘤生长的影响。方法体外培养鼠源性C6胶质瘤细胞,转染miR-103a-3p mimics质粒过表达miR-103a-3p,转染miR-103a-3p mimics+pcDNA-PDK4质粒分析PDK4过表达对miR-103a-3p过表达的影响;EDU法检测细胞增殖活性;qRT-PCR检测miR-103a-3p、PDK4 mRNA表达水平;流式细胞仪检测细胞凋亡率;Transwell实验检测细胞侵袭能力;免疫印迹法检测E-cadherin、N-cadherin、vimentin蛋白表达水平。取40只裸鼠,其中20只皮下注射未转染质粒的C6细胞、20只皮下注射转染miR-103a-3p mimics质粒的C6细胞构建移植瘤模型,分析miR-103a-3p mimics过表达对移植瘤生长的影响。结果生物信息学及双荧光素酶试验证实PDK4是miR103a-3p的靶点。过表达miR-103a-3p明显降低C6细胞PDK4、Ki67、PCNA、N-cadherin、vimentin表达水平(P<0.05),明显抑制C6细胞增殖活性、侵袭能力(P<0.05),明显增加C6细胞E-cadherin表达水平、细胞凋亡率(P<0.05)。过表达PDK4明显抑制过表达miR-103a-3p对C6胶质瘤细胞的作用(P<0.05)。过表达miR-103a-3p明显抑制裸鼠移植瘤生长(P<0.05),抑制肿瘤组织PDK4、Ki67、vimentin表达(P<0.05)。结论过表达miR-103a-3p通过靶向抑制PDK4表达,一方面抑制胶质瘤细胞增殖、促进胶质瘤细胞凋亡,从而抑制胶质瘤生长;另一方抑制胶质瘤上皮-间质转化过程,从而抑制胶质瘤细胞侵袭。
Objective To explore the effect of miR-103a-3p overexpression on the malignant biological behavior of glioma C6 cells and the growth of transplanted glioma in nude mice.Methods Murine C6 glioma cells were cultured in vitro,miR-103a-3p mimics plasmid was transfected into the C6 cells to overexpress the miR-103a-3p,miR-103a-3p mimics+pcDNA-PDK4 plasmid was transfected into the C6 cells to analyze the effect of PDK4 overexpression on miR-103a-3p overexpression.C6 glioma cells were subcutaneously injected into 40 nude mice to construct a transplanted tumor model,of which 20 received subcutaneous injection of C6 cells without plasmid transfection and 20 transfected miR-103a-3p mimics plasmid C6 cells.EDU method was used to detect the cell proliferation activity.qRT-PCR was used to detect the miR-103a-3p and PDK4 mRNA expression levels.Flow cytometry was used to detect cell apoptosis rate.Transwell experiment was used to detect cell invasion ability.Western blotting was used to detect the protein expression levels of E-cadherin,N-cadherin,and vimentin.Results Bioinformatic analysis and dual luciferase test confirmed that PDK4 was the direct target of miR-103a-3p.Overexpression of miR-103a-3p significantly reduced the expression levels of PDK4,Ki67,PCNA,N-cadherin,and vimentin in C6 cells(P<0.05),significantly inhibited C6 cell proliferation activity and invasion ability(P<0.05),and significantly increased C6 cell E-cadherin expression level(P<0.05),significantly increased the apoptosis rate of C6 cells(P<0.05).Overexpression of PDK4 significantly inhibited the effect of overexpression of miR-103a-3p on C6 glioma cells(P<0.05).Overexpression of miR-103a-3p significantly inhibited the growth of transplanted tumors in nude mice(P<0.05),and inhibited the expression of PDK4,Ki67,and vimentin in tumor tissues(P<0.05).Conclusions Overexpression of miR-103a-3p can inhibit glioma cell proliferation and promote glioma cell apoptosis through targeted inhibition of PDK4 expression,thereby inhibiting glioma growth;on the other hand
作者
郭放
万学锋
刘献志
杨强
李太平
GUO Fang;WAN Xue-feng;LIU Xian-zhi;YANG Qiang;LI Tai-ping(Department of Neurosurgery,Zhengzhou Yihe Hospital,Zhengzhou 450003,China;Department of Neurosurgery,Zhumadian Central Hospital Affiliated to Huanghuai University,Zhumadian 463000,China;Department of Neurosurgery,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450003,China)
出处
《中国临床神经外科杂志》
2021年第6期442-448,共7页
Chinese Journal of Clinical Neurosurgery
基金
2016年河南省攻关计划项目(201604213)。
