摘要
目的探索miR-130a-3p对胶质母细胞瘤(glioblastoma,GBM)替莫唑胺(temozolomide,TMZ)化疗敏感性的影响与机制。方法采用实时荧光逆转录定量聚合酶链反应(real-time reverse transcription quantitative polymerase chain reaction,RT-qPCR)检测胶质瘤细胞T98G细胞和TMZ耐药T98G-R细胞中miR-130a-3p以及胞质多聚腺苷酸化元件结合蛋白4(cytoplasmic polyadenylation element binding protein 4,CPEB4)的相对表达水平。四甲基偶氮唑盐(methylthiazolyldiphenyl-tetrazolium bromide,MTT)实验检测胶质瘤细胞的增殖能力,流式细胞术检测细胞凋亡情况,荧光素酶报告实验检测miR-130a-3p与CPEB4的结合。结果T98G-R耐药细胞中miR-130a-3p表达水平较T98G细胞降低(0.25±0.08 vs.1.00±0.05,P<0.01),CPEB4表达水平较T98G细胞升高(1.37±0.17 vs.1.00±0.05,P<0.01)。MTT实验及流式细胞术结果表明,miR-130a-3p过表达可抑制胶质瘤细胞的增殖,促进TMZ诱导的细胞凋亡,但其可以被CPEB4过表达所逆转。荧光素酶报告实验显示,miR-130a-3p可以与CPEB4的3′-非翻译区结合,过表达miR-130a-3p抑制CPEB4的表达。结论miR-130a-3p通过靶向抑制CPEB4表达,促进胶质瘤细胞凋亡,增加GBM对TMZ的敏感性。
Objective To explore the effect and mechanism of miR-130 a-3 p on chemosensitivity of temozolomide(TMZ)in glioblastoma(GBM).Methods The relative expression levels of miR-130 a-3 p and cytoplasmic polyadenylation element binding protein 4(CPEB4)in T98 G cells and TMZ resistant T98 G-R cells were detected by real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR).The proliferation ability of glioma cells was detected by methylthiazolyldiphenyl-tetrazolium bromide(MTT),and the flow cytometry was applied to detect cell apoptosis.Luciferase report experiment verified the combination of miR-130 a-3 p and CPEB4.Results The expression level of miR-130 a-3 p in T98 G-R TMZ resistant cells was lower than that in T98 G cells(0.25±0.08 vs.1.00±0.05,P<0.01);the expression level of CPEB4 was higher than that in T98 G cells(1.37±0.17 vs.1.00±0.05,P<0.01).MTT and flow cytometry showed that miR-130 a-3 p overexpression could inhibit the proliferation of glioma cells and promote temozolomide induced apoptosis,whereas forced expression of CPEB4 partially could abolish this effect.Luciferase report experiment showed that miR-130 a-3 p could bind to the 3′-untranslated region of CPEB4,and overexpression of miR-130 a-3 p could inhibit the expression of CPEB4.Conclusion miR-130 a-3 p can promote cell apoptosis and increase the sensitivity of GBM to TMZ by targeted inhibition of CPEB4 expression.
作者
魏志豪
刘洪超
史康克
张雨
李佳琼
WEI Zhihao;LIU Hongchao;SHI Kangke;ZHANG Yu;LI Jiaqiong(Department of Pathology,the Yiluo Hospital of Luoyang,Luoyang Henan 471023,China)
出处
《华南国防医学杂志》
CAS
2022年第9期683-688,共6页
Military Medical Journal of South China
基金
洛阳市医学重点培育学科项目(洛卫科教2022-5号)。