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微小RNA-200c和微小RNA-141协同抑制锌指E盒结合蛋白1,并抑制胶质瘤细胞的生长和迁移。

MiR-200c and miR-141 inhibit ZEB1 synergistically and suppress glioma cell growth and migration.

作者信息

Guo E, Wang Z, Wang S

机构信息

Department of Neurosurgery, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.

出版信息

Eur Rev Med Pharmacol Sci. 2016 Aug;20(16):3385-91.

Abstract

OBJECTIVE

This study aimed to investigate the expression of miR-200c and miR-141 in glioma tissues and cell lines and then to study their regulative effect on ZEB1 expression and on glioma cell growth and migration.

MATERIALS AND METHODS

QRT-PCR analysis was performed to detect miR-200c and miR-141 expression in 10 paired glioma tissues and adjacent normal tissues from patients with glioblastoma multiforme (GBM) and in glioma cell lines. U87 and U251 cells were transfected with miR-200c mimics, miR-141 mimics or ZEB1 siRNA respectively. ZEB1 expression was detected qRT-PCR and Western blot assay. MTT assay, flow cytometry and wound healing assay were performed to examine the tumor suppressive effects of the miR-200c/miR-141-ZEB1 axis on glioma cells.

RESULTS

Both miR-200c and miR-141 were significantly lower in glioma tissues than in adjacent normal tissues. The glioma cell lines, including U87, U251 and A172 also had significantly decreased miR-200c and miR-141 expression than normal tissues. ZEB1 expression had at least two-fold increase in glioma tissues than in normal tissues. Both miR-200c and miR-141 could significantly induce ZEB1 mRNA degradation and suppress ZEB1 protein expression. ZEB1 siRNA presented similar growth and migration inhibiting and apoptosis inducing effect to miR-200c and miR-141 mimics in U87 cells.

CONCLUSIONS

MiR-200c and miR-141 are significantly downregulated in glioma tissues and cell lines and can significantly induce ZEB1 mRNA degradation and suppress ZEB1 protein expression in the cells. ZEB1 is a functional downstream target of miR-200c and miR-141 in inhibiting glioma cell growth and migration.

摘要

目的

本研究旨在调查miR - 200c和miR - 141在胶质瘤组织和细胞系中的表达情况,进而研究它们对ZEB1表达以及胶质瘤细胞生长和迁移的调控作用。

材料与方法

采用实时定量聚合酶链反应(QRT - PCR)分析检测10对多形性胶质母细胞瘤(GBM)患者的胶质瘤组织及相邻正常组织以及胶质瘤细胞系中miR - 200c和miR - 141的表达。分别用miR - 200c模拟物、miR - 141模拟物或ZEB1小干扰RNA(siRNA)转染U87和U251细胞。通过qRT - PCR和蛋白质免疫印迹法检测ZEB1的表达。采用MTT法、流式细胞术和伤口愈合试验检测miR - 200c/miR - 141 - ZEB1轴对胶质瘤细胞的肿瘤抑制作用。

结果

miR - 200c和miR - 141在胶质瘤组织中的表达均显著低于相邻正常组织。包括U87、U251和A172在内的胶质瘤细胞系中miR - 200c和miR - 141的表达也明显低于正常组织。ZEB1在胶质瘤组织中的表达比正常组织至少增加两倍。miR - 200c和miR - 141均可显著诱导ZEB1信使核糖核酸(mRNA)降解并抑制ZEB1蛋白表达。在U87细胞中,ZEB1 siRNA对细胞生长和迁移的抑制作用以及诱导凋亡的作用与miR - 200c和miR - 141模拟物相似。

结论

miR - 200c和miR - 141在胶质瘤组织和细胞系中显著下调,可显著诱导细胞中ZEB1 mRNA降解并抑制ZEB1蛋白表达。在抑制胶质瘤细胞生长和迁移方面,ZEB1是miR - 200c和miR - 141的功能性下游靶点。

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