miR-138-5p调控CDC5L表达对胶质瘤细胞增殖、侵袭的影响

Influences of miR-138-5p on proliferation and invasion of glioma cells by regulating CDC5L expression

目的探讨微RNA(microRNA,miR)-138-5p对胶质瘤细胞增殖、侵袭的调控机制。方法体外培养人胶质瘤U251细胞,采用随机数字表法将其分为对照组(n=6)、miR-NC组(n=6)、miR-138-5p组(n=6)、miR-138-5p+pc-NC组(n=6)、miR-138-5p+pc-细胞分裂周期5样(cell division cycle 5-like,CDC5L)组(n=6)。采用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)检测细胞中miR-138-5p、CDC5L mRNA水平,检测细胞增殖活力、细胞周期和细胞凋亡以及细胞侵袭能力,检测细胞中CDC5L和侵袭相关蛋白表达。结果miR-138-5p的过表达可下调CDC5L mRNA和蛋白水平,降低胶质瘤细胞增殖活力和侵袭能力,并诱导G2/M期细胞周期停滞,促进细胞凋亡,降低N-钙粘蛋白、波形蛋白水平,升高E-钙粘蛋白水平,差异均有统计学意义(P<0.05);上调CDC5L表达可明显逆转过表达miR-138-5p对胶质瘤细胞恶性生物学行为的影响(P<0.05);与miR-NC和CDC5L-WT质粒共转染比较,miR-138-5p mimic和CDC5L-WT质粒共转染的细胞中荧光素酶活性降低,差异有统计学意义(P<0.05)。结论过表达miR-138-5p可能通过靶向下调CDC5L,抑制胶质瘤细胞增殖和侵袭。

Objective To investigate the regulatory mechanism of microRNA-138-5p(miR-138-5p)on the proliferation and invasion of glioma cells.Methods Human glioma U251 cells were cultured in vitro and divided into control group(n=6),miR-NC group(n=6),miR-138-5p group(n=6),miR-138-5p+pc-NC group(n=6),and miR-138-5p+pc-cell division cycle 5-like(CDC5L)group(n=6)according to the random number table method.Real-time quantitative PCR(RT-qPCR)was performed to measure the levels of miR-138-5p and CDC5L mRNA in cells,to measure cell proliferation viability,cell cycle and apoptosis,and cell invasion ability,and to measure the expression of CDC5L and invasion-related proteins in cells.Results Overexpression of miR-138-5p was able to down-regulate CDC5L mRNA and protein levels,reduce the proliferative viability and invasive ability of glioma cells,induce cell cycle arrest in G2/M phase,promote cell apoptosis,and reduce N-cadherin and Vimentin levels,increased E-cadherin level,and the differences were statistically significant(P<0.05);up-regulation of CDC5L expression was able to significantly reverse the effects of overexpression of miR-138-5p on the malignant biological behavior of of glioma cell(P<0.05);the dual luciferase assay showed that compared with co-transfection of miR-NC and CDC5L-WT plasmids,the luciferase activity in cells co-transfected with miR-138-5p mimic and CDC5L-WT plasmids was significantly decreased(P<0.05).Conclusion Overexpression of miR-138-5p may inhibit the proliferation and invasion of glioma cells by targeting down-regulation of CDC5L.