miR-34a-5p 通过靶向 HMGA2 抑制胶质瘤的发生发展并增强替莫唑胺诱导的胶质瘤细胞毒性。

MicroRNA-34a-5p suppresses tumorigenesis and progression of glioma and potentiates Temozolomide-induced cytotoxicity for glioma cells by targeting HMGA2.

机构信息

Department One of Neurosurgery, Zhoukou Central Hospital, Zhoukou, Henan, China.

出版信息

Eur J Pharmacol. 2019 Jun 5;852:42-50. doi: 10.1016/j.ejphar.2019.03.005. Epub 2019 Mar 6.

DOI:10.1016/j.ejphar.2019.03.005

Abstract

Glioma is a frequently diagnosed brain tumors and Temozolomide (TMZ) is a common chemotherapeutic drug for glioma. High mobility group AT-hook 2 (HMGA2) was reported to be linked with glioma pathogenesis and Temozolomide (TMZ)-induced cytotoxicity. Our present study aimed to further search for the upstream regulatory microRNAs (miRNAs) of HMGA2 in glioma. RT-qPCR assay was conducted to measure the expression of HMGA2 mRNA and microRNA-34a-5p (miR-34a-5p). HMGA2 protein expression was examined by western blot assay. Cell proliferative ability and cell viability was assessed by CCK-8 assay. Cell migratory and invasive capacities were estimated by Transwell migration and invasion assay. Bioinformatics analysis and luciferase reporter assay was conducted to investigate the potential interaction between miR-34a-5p and HMGA2. Mouse xenograft experiments were performed to further test the roles of TMZ, miR-34a-5p and HMGA2, alone or in combination, in glioma tumorigenesis in vivo. We found HMGA2 expression was notably upregulated in glioma tissues and cells, and associated with glioma grade and poor prognosis. HMGA2 knockdown or miR-34a-5p overexpression inhibited migration, invasion, proliferation and enhanced TMZ-induced cytotoxicity in glioma cells. Moreover, HMGA2 was a target of miR-34a-5p. And, miR-34a-5p expression was remarkably reduced in glioma tissues and cells. MiR-34a-5p exerted its function through targeting HMGA2 in glioma cells. HMGA2 knockdown or miR-34a-5p overexpression inhibited tumor growth and enhanced TMZ-mediated anti-tumor effect in glioma xenograft models. We concluded MiR-34a-5p suppressed tumorigenesis and progression of glioma and potentiated TMZ-induced cytotoxicity for glioma cells by targeting HMGA2, deepening our understanding on molecular basis of HMGA2 in glioma.

摘要

神经胶质瘤是一种常见的脑肿瘤，替莫唑胺（TMZ）是治疗神经胶质瘤的常用化疗药物。高迁移率族 AT 盒 2（HMGA2）与神经胶质瘤的发病机制和替莫唑胺（TMZ）诱导的细胞毒性有关。本研究旨在进一步寻找 HMGA2 在神经胶质瘤中的上游调节 microRNA（miRNA）。通过 RT-qPCR 检测 HMGA2mRNA 和 microRNA-34a-5p（miR-34a-5p）的表达。通过 Western blot 检测 HMGA2 蛋白表达。通过 CCK-8 检测细胞增殖能力和细胞活力。通过 Transwell 迁移和侵袭实验评估细胞迁移和侵袭能力。通过生物信息学分析和荧光素酶报告基因实验研究 miR-34a-5p 与 HMGA2 之间的潜在相互作用。进行小鼠异种移植实验进一步测试 TMZ、miR-34a-5p 和 HMGA2 单独或联合在体内对神经胶质瘤发生的作用。我们发现 HMGA2 在神经胶质瘤组织和细胞中表达明显上调，并与神经胶质瘤分级和预后不良相关。HMGA2 敲低或 miR-34a-5p 过表达抑制神经胶质瘤细胞的迁移、侵袭、增殖，并增强 TMZ 诱导的细胞毒性。此外，HMGA2 是 miR-34a-5p 的靶标。而且，miR-34a-5p 在神经胶质瘤组织和细胞中的表达明显降低。miR-34a-5p 通过靶向 HMGA2 在神经胶质瘤细胞中发挥作用。HMGA2 敲低或 miR-34a-5p 过表达抑制神经胶质瘤异种移植模型中的肿瘤生长并增强 TMZ 介导的抗肿瘤作用。我们得出结论，miR-34a-5p 通过靶向 HMGA2 抑制神经胶质瘤的发生和进展，并增强 TMZ 诱导的神经胶质瘤细胞毒性，加深了我们对 HMGA2 在神经胶质瘤中的分子基础的理解。