miR-23c靶向调控MTDH抑制胶质瘤U87细胞增殖迁移与侵袭

miR-23c inhibits the proliferation,migration,and invasion of glioma U87 cells via targeted regulation of MTDH

目的探讨miR-23c对胶质瘤细胞U87增殖、迁移和侵袭的影响及初步机制。方法将miR-23c mimics转染于U87细胞中,用miR-无关序列转染于U87细胞做为Control组,采用MTT法、细胞划痕、Transwell侵袭实验观察miR-23 c对U87细胞增殖、迁移和侵袭的影响;采用生物信息学软件分析miR-23c潜在的靶基因;采用Western blot、RT-PCR和萤光素酶报告基因检测miR-23c对靶基因的调控作用;在转染miR-23c mimics的基础上同时转染MTDH质粒,通过MTT法、细胞划痕、transwell侵袭实验观察转染MTDH对miR-23c抑制U87细胞增殖、迁移和侵袭的影响。结果 MTT实验显示,miR-23 c组U87细胞的OD值为(0. 668±0. 032),明显低于Control组的(1. 031±0. 060)(P <0. 01);细胞划痕实验显示,miR-23c组细胞划痕愈合率(0. 35±0. 02)明显低于对照组的(0. 59±0. 03)(P <0. 01);Transwell侵袭实验显示,miR-23 c组U87细胞穿过基质胶的细胞数(153. 2±8. 30)明显低于与对照组的(348. 4±12. 12)(P <0. 01);Western blot、RT-PCR和萤光素酶报告基因检测表明MTDH是miR-23 c直接调控的靶基因;MTT实验显示,miR-23 c+MTDH组U87细胞的OD值为(1. 025±0. 059),明显高于miR-23 c组的(0. 672±0. 024)(P <0. 01);细胞划痕实验显示,miR-23 c+MTDH组细胞划痕愈合率为(0. 45±0. 04),明显高于miR-23 c组的(0. 31±0. 03)(P <0. 05);Transwell侵袭实验显示,miR-23c+MTDH组U87细胞穿过基质胶的细胞数(260. 9±10. 23),明显高于miR-23c组的(148. 4±9. 4)(P <0. 01)。结论上调miR-23 c能明显抑制胶质瘤细胞的增殖、迁移和侵袭能力,其机制可能与其下调MTDH表达有关。

Objective To investigate the effect of miR-23 c on the proliferation,migration,and invasion of glioma U87 cells and preliminary mechanisms. Methods U87 cells were transfected with miR-23 c mimics,and the U87 cells transfected with miR-negative control were established as control group. MTT assay,wound healing assay,and Transwell invasion experiment were used to observe the effect of miR-23 c on the proliferation,migration,and invasion of U87 cells. Bioinformatics software was used to analyze the potential target genes of miR-23 c,and Western blot,RT-PCR,and luciferase reporter gene assay were used to observe the regulatory effect of miR-23 c on target genes. After U87 cells were co-transfected with miR-23 c mimics and MTDH,MTT assay,wound healing assay,and Transwell invasion experiment were used to analyze the effect of MTDH transfection on the proliferation,migration,and invasion of U87 cells inhibited by miR-23 c. Results The MTT assay showed that the miR-23 c group had a significantly lower OD value of U87 cells than the control group( 0. 668 ± 0. 032 vs 1. 031 ± 0. 060,P 0. 01);the wound healing assay showed that the miR-23 c group had a significantly lower wound healing rate than the control group( 0. 35 ± 0. 02 vs 0. 59 ± 0. 03,P 0. 01);the Transwell invasion experiment showed that the miR-23 c group had a significantly lower number of U87 cells passing through the matrigel than the control group( 153. 2 ± 8. 30 vs 348. 4 ± 12. 12,P 0. 01). Western blot,RT-PCR,and luciferase reporter gene assay showed that MTDH was a target gene directly regulated by miR-23 c. The MTT assay showed that the miR-23 c + MTDH group had a significantly higher OD value of U87 cells than the miR-23 c group( 1. 025 ± 0. 059 vs 0. 672 ± 0. 024,P 0. 01);the wound healing assay showed that the miR-23 c + MTDH group had a significantly higher wound healing rate of U87 cells than the miR-23 c group( 0. 45 ± 0. 04 vs 0. 31 ±0. 03,P 0. 05);the Transwell invasion experiment showed that the miR-23 c + MTDH group had a significantly higher number of U87 cells passing through the matrigel than the miR-23 c group( 260. 9 ± 10. 23 vs 148. 4 ± 9. 4,P 0. 01). Conclusions Upregulation of miR-23 c can significantly inhibit the proliferation,migration,and invasion of glioma cells,possibly by downregulating the expression of MTDH.