微小RNA⁃630对U251神经胶质瘤细胞增殖和凋亡的影响  

Effects of microRNA-630 on proliferation and apoptosis of U251 glioma cells

作　　者：钟曌 孙世辉 祝刚 郭昊 ZHONG Zhao;SUN Shihui;ZHU Gang;GUO Hao(Department of Neurology,the Second Affiliated Hospital of Xi'an Air Force Military Medical University,Xi'an 710038,China)

机构地区：[1]空军军医大学附属第二医院神经内科,西安710038 [2]空军军医大学附属第二医院神经外科,710038

出　　处：《临床肿瘤学杂志》2023年第4期326-331,共6页Chinese Clinical Oncology

基　　金：国家自然科学基金资助项目(81903072)。

摘　　要：目的探讨微小RNA-630(miR-630)对U251神经胶质瘤细胞增殖和凋亡的影响及其调控机制。方法荧光定量PCR(qPCR)检测人脑胶质瘤组织和细胞的miR-630表达水平。采用脂质体转染法向U251细胞转染miR-630模拟物mimics(miR-630 mimics组)及阴性对照序列(miR-NC mimics组),另选取未转染细胞为空白对照(Control)组,经qPCR验证miR-630 mimics显著增强miR-630基因表达后进一步进行功能研究:CCK-8和Hoechst染色法检测细胞增殖变化,采用流式细胞术、qPCR和Western blot分别检测凋亡细胞百分率和凋亡基因表达,qPCR和双荧光素酶报告基因系统分别检测20S蛋白酶体α1亚基(PSMA1)的表达及其与miR-630靶向关系验证。结果miR-630在胶质瘤组织和细胞表达下调。与Control组和miR-NC mimics组相比,miR-630 mimics组U251细胞增殖能力下降,细胞凋亡增加,抗凋亡基因Bcl-2表达降低,cleaved caspase-3表达增加。过表达miR-630显著降低PSMA1表达和荧光素酶活性,而对突变后PSMA1荧光素酶活性无影响。PSMA1在胶质瘤组织表达上调,且其表达与miR-630呈负相关。结论过表达miR-630可能通过降低PSMA1表达来抑制U251神经胶质瘤细胞增殖并促进凋亡。Objective To investigate the effects of microRNA-630(miR-630)on proliferation and apoptosis of human glioma U251 cells and potential mechanism.Methods Real-time fluorescence quantitative PCR(qPCR)was applied to detect miR-630 expression in glioma tissues and cell lines.U251 cells were transfected with miR-630 mimics(miR-630 mimics group)and negative control sequence(miR-NC mimics group)using liposomes,and untransfected cells were selected as control group.MiR-630 mimics significantly enhanced the expression of miR-630 as verified by qPCR,and further functional research were conducted.The cell proliferation were detected by CCK-8 and Hoechst staining.The apoptotic rate and genes expression were determined by flow cytometry,qPCR and Western blot.QPCR and dual fluorescence reporting system were used to detect the expression and enzyme activity of proteasome 20S subunit alpha 1(PSMA1).Results MiR-630 was weakly expressed in glioma tissues and cell lines.Compared to the control group and miR-NC mimics group,there were blunted cell proliferation,enhanced cell apoptosis,decreased anti-apoptotic gene Bcl-2 and increased cleaved caspase-3 in the miR-630 mimics group.Overexpression of miR-630 significantly reduced PSMA1 expression and luciferase activity(P<0.05),while the luciferase activity was uninfluenced after mutation of PSMA1.PSMA1 was highly expressed in glioma tissues and negatively correlated with miR-630 expression.Conclusion Overexpression of miR-630 can inhibit proliferation and promote apoptosis of U251 glioma cells by decreasing PSMA1 expression.

关 键 词：神经胶质瘤 微小RNA-630 增殖 凋亡 20S蛋白酶体α1亚基 

分 类 号：R739.41[医药卫生—肿瘤]