MicroRNA-105通过靶向调控SOX9的表达抑制脑胶质瘤进展

发布时间：2017-12-31 19:11

本文关键词：MicroRNA-105通过靶向调控SOX9的表达抑制脑胶质瘤进展　出处：《南方医科大学》2017年博士论文　论文类型：学位论文

更多相关文章： miR-105 SOX9 TCF4 脑胶质瘤 增殖凋亡

【摘要】：脑胶质瘤是最常见的原发性脑肿瘤,占所有颅内肿瘤的46%左右,每年大约有35万人被诊断出患有胶质瘤。虽然目前胶质瘤的治疗取得了很大进展,但是胶质母细胞瘤中位生存期仅有15个月。因此,迫切需要新的分子靶点和处理手段。microRNAs(miRNAs)是近年来发现的一类非编码小分子RNA。据报道,miRNAs参与了生命过程中一系列的重要进程,包括发育调控、器官形成、细胞增殖和凋亡。最近研究表明,许多miRNAs在肿瘤中异常表达,并参与肿瘤的发生和发展,已成为当前肿瘤诊断和治疗研究的热点之一。其中,miR-105在多种肿瘤中被证明表达异常,发挥抑癌基因或癌基因的作用。同时,研究表明miR-105在脑胶质瘤组织中低表达,且与不良预后相关。但是,miR-105在脑胶质瘤发生和发展中的作用和调控机制还不清楚。本研究采用qRT-PCR、荧光素酶报告实验、CCK-8实验、细胞流式和transwell小室检测等方法,研究miR-105在脑胶质瘤组织和细胞中的表达,检测miR-105对脑胶质瘤细胞增殖、凋亡和侵袭的影响,筛选miR-105的直接下游靶基因,进而探讨miR-105可能的调控机制,为后续在脑胶质瘤治疗中的应用提供一定的理论依据。结果表明:1)miR-105在脑胶质瘤组织和细胞中均低表达。2)miR-105上调可以抑制脑胶质瘤细胞增殖、侵袭并诱导凋亡。3)荧光素酶报告实验证明SOX9是miR-105的直接下游靶基因,并且SOX9在脑胶质瘤组织和细胞中高表达,与miR-105的表达呈负相关。4)SOX9干扰抑制脑胶质瘤细胞增殖、侵袭并诱导凋亡。5)SOX9过表达可以回复miR-105上调对脑胶质瘤细胞增殖、侵袭的抑制和对凋亡的诱导影响。6)SOX9 干扰可以抑制 SOX9、TCF4、c-MYC、cyclin D1 和 AXIN2 蛋白的表达,TCF4过表达可以回复SOX9干扰对SOX9、TCF4、c-MYC、cyclinD1和AXIN2蛋白表达的抑制作用,并且可以回复SOX9干扰对脑胶质瘤细胞增殖、侵袭的抑制和对凋亡的诱导影响。7)miR-105 上调抑制 SOX9、TCF4、c-MYC、cyclin D1 和 AXIN2 蛋白的表达,SOX9过表达可以回复miR-105上调对SOX9、TCF4、c-MYC、cyclinD1和AXIN2蛋白表达的抑制作用,TCF4过表达也可以回复miR-105 上调对 TCF4、c-MYC、cyclin D1 和 AXIN2 蛋白表达的抑制作用,并且TCF4过表达可以回复miR-105上调对脑胶质瘤细胞增殖、侵袭的抑制和对凋亡的诱导影响。综上所述,miR-105在脑胶质瘤组织和细胞中低表达,并且通过靶向SOX9/TCF4信号通路抑制脑胶质瘤细胞增殖和侵袭并诱导凋亡。
[Abstract]:Gliomas are the most common primary brain tumors, accounting for about 46% of all intracranial tumors, and about 350,000 people are diagnosed with gliomas each year, although much progress has been made in the treatment of gliomas. But the median survival time of glioblastoma is only 15 months. There is an urgent need for new molecular targets and treatment methods. MicroRNAs-miRNAsis a kind of non-coding small molecule RNA. reported in recent years. MiRNAs is involved in a series of important processes in life, including developmental regulation, organ formation, cell proliferation and apoptosis. Recent studies have shown that there are many abnormal expressions of miRNAs in tumors. And participate in the occurrence and development of tumors, has become one of the hot spots in the diagnosis and treatment of cancer, among which the expression of miR-105 has been proved to be abnormal in many kinds of tumors. At the same time, studies have shown that miR-105 is low expressed in gliomas and is associated with poor prognosis. The role and regulatory mechanism of miR-105 in the genesis and development of glioma is unclear. In this study, qRT-PCR, luciferase report experiment and CCK-8 were used. The expression of miR-105 in glioma tissues and cells was studied by flow cytometry and transwell chamber detection. The proliferation of glioma cells was detected by miR-105. The direct downstream target gene of miR-105 was screened by the effect of apoptosis and invasion, and the possible regulation mechanism of miR-105 was discussed. The results show that the expression of 1: 1 miR-105 is low in glioma tissues and glioma cells. Upregulation of miR-105 can inhibit the proliferation of glioma cells. Luciferase report showed that SOX9 is a direct downstream target gene of miR-105 and that SOX9 is highly expressed in glioma tissues and cells. There was a negative correlation with the expression of miR-105. The interference of SOX9 inhibited the proliferation of glioma cells. The overexpression of SOX9 in invading and inducing apoptosis could reverse the proliferation of glioma cells induced by miR-105. Inhibition of invasion and induction of apoptosis. The interference of SOX9 can inhibit the expression of cyclin D1 and AXIN2 protein. The overexpression of TCF4 could restore the inhibitory effect of SOX9 interference on the expression of CyclinD1 and AXIN2 proteins in SOX9 TCF4 and c-MYCU. And the inhibition of SOX9 interference on the proliferation, invasion and apoptosis of glioma cells. 7% miR-105 up-regulated the inhibition of SOX9 TCF4 c-MYC. The overexpression of cyclin D1 and AXIN2 protein (SSOX9) could respond to the up-regulation of miR-105 on SOX9 TCF4 c-MYC. The inhibitory effect of cyclinD1 and AXIN2 protein expression and the overexpression of TCF4 could also reverse the up-regulation of TCF4 c-MYC by miR-105. The expression of cyclin D1 and AXIN2 protein was inhibited, and TCF4 overexpression could up-regulate the proliferation of glioma cells by miR-105. The inhibition of invasion and the induction of apoptosis. The proliferation and invasion of glioma cells were inhibited and apoptosis induced by targeting SOX9/TCF4 signaling pathway.
【学位授予单位】：南方医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R739.41

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