MiR-92bⅠ抑制胶质瘤恶性表型体外研究
发布时间：2018-10-16 22:34
【摘要】：背景和目的： 脑胶质瘤是中枢神经系统最常见的原发性肿瘤,由于恶性胶质瘤有侵袭性生长、增殖速度快等特征,应用目前的治疗措施如外科手术切除、术后放疗、化疗和免疫治疗等不能治愈,患者复发率很高,临床预后甚差。造成这种现状的原因,很大部位是对脑胶质瘤的病因,发病机制的认识不充分。为此,必须寻找与胶质瘤发生、发展相关的基因和信号通路,进一步深入了解胶质瘤发生发展的分子病理机制,寻找胶质瘤治疗的新靶点,在此基础上开拓胶质瘤治疗的新策略和新手段。 有研究证实,Nemo-like kinase(NLK)是一种抑癌基因,在体外的恶性肿瘤细胞株中的研究发现它可以通过促进肿瘤细胞的凋亡而起到抑癌作用,研究表明NLK能够促进人的结肠癌细胞DLD-1细胞株,乳腺癌细胞株和胶质瘤细胞株的凋亡[1-3]。但NLK在脑胶质瘤中的研究甚少,故可以初步认定NLK是一个胶质瘤抑癌基因。此外,为了寻找可调控NLK表达的microRNA,我们搜索了预测microRNA靶基因网站,包括TARGETSCAN, MIRANDA等,发现有很多的microRNA,包括niR-92b在NLK mRNA3'UTR上有潜在的结合位点,故本课题重点研究了miR-92b I抑制胶质瘤增殖、细胞周期进展、凋亡和侵袭的影响及其可能的作用机制。 方法和结果： 本课题研究分为以下两个部分： 第一部分中应用Real time PCR的方法,检测了5个胶质瘤细胞系和38例不同级别人脑胶质瘤组织中miR-92b的表达水平。结果表明miR-92b在胶质瘤组织和细胞系中的表达较正常组织上调,并与肿瘤级别呈正相关。 在课题第二部分,miR-92b Ⅰ抑制胶质瘤恶性表型的体外研究。将niR-92b Ⅰ转染至人脑胶质瘤细胞系SNB19和LN229中。采用Real time PCR检测转染后胶质瘤细胞的miR-92b表达水平以鉴定抑制效果；四甲基偶氮唑蓝(MTT)实验评价细胞的增殖率；流式细胞术检测细胞周期变化；Annexin Ⅴ法检测细胞早期凋亡;Transwell实验检测细胞侵袭能力变化。Real time PCR检测结果显示转染miR-92b Ⅰ后,肿瘤细胞miR-92b表达下降,miR-92b Ⅰ组细胞增殖活性降低,细胞侵袭能力明显受到抑制,细胞周期S期缩短,阻滞在G0/G1期以及早期凋亡增加。该实验说明miR-92b是癌微RNA(OncomiRs),可能成为胶质瘤治疗的潜在靶点。 结论： miR-92b在胶质瘤中表达上调,低级别肿瘤与高级别肿瘤比较,其差异具有显著性,但是正常组织与低级别肿瘤比较,差异没有显著性。转染miR-92b Ⅰ至人胶质瘤细胞系,可抑制胶质瘤细胞的增殖活性和侵袭能力,细胞周期S期缩短,阻滞在G0/G1期,并促进细胞凋亡,由此,初步认定miR-92b为癌微RNA,可成为胶质瘤基因治疗的候选靶标。这是首次有关miR-92b在人脑胶质瘤中的表达以及对胶质瘤恶性表型的影响的研究报道,为进一步认识脑胶质瘤的分子病理机制及新一类的靶向治疗提供实验依据。
[Abstract]:Background and objective: glioma is the most common primary tumor in the central nervous system. Chemotherapy and immunotherapy can not be cured, the recurrence rate is very high, the clinical prognosis is very poor. Most of the causes of this situation are the etiology of glioma, and the pathogenesis of glioma is not well understood. Therefore, it is necessary to search for genes and signal pathways related to glioma development, to further understand the molecular pathological mechanism of glioma development, and to find new targets for the treatment of glioma. On this basis, a new strategy and a new method for the treatment of glioma were developed. Some studies have confirmed that Nemo-like kinase (NLK) is a tumor suppressor gene. In vitro, it has been found that Nemo-like kinase (NLK) can inhibit cancer by promoting apoptosis of tumor cells. It has been shown that NLK can promote human colon cancer cell DLD-1 cell line. Apoptosis of breast cancer cell line and glioma cell line [1-3]. However, there are few studies of NLK in gliomas, so it can be preliminarily concluded that NLK is a tumor suppressor gene. In addition, in order to find microRNA, that can regulate the expression of NLK, we searched microRNA target gene websites, including TARGETSCAN, MIRANDA, and found that many microRNA, including niR-92b, had potential binding sites on NLK mRNA3'UTR. Therefore, this study focused on the inhibition of glioma proliferation by miR-92b I. Cell cycle progression, the effects of apoptosis and invasion and their possible mechanisms. Methods and results: the study was divided into the following two parts: in the first part, the expression of miR-92b in 5 glioma cell lines and 38 cases of human gliomas of different grades were detected by the method of Real time PCR. The results showed that the expression of miR-92b in glioma tissues and cell lines was higher than that in normal tissues, and there was a positive correlation between the expression of miR-92b and tumor grade. In the second part of the study, miR-92b 鈪，
本文编号：2275836
[Abstract]:Background and objective: glioma is the most common primary tumor in the central nervous system. Chemotherapy and immunotherapy can not be cured, the recurrence rate is very high, the clinical prognosis is very poor. Most of the causes of this situation are the etiology of glioma, and the pathogenesis of glioma is not well understood. Therefore, it is necessary to search for genes and signal pathways related to glioma development, to further understand the molecular pathological mechanism of glioma development, and to find new targets for the treatment of glioma. On this basis, a new strategy and a new method for the treatment of glioma were developed. Some studies have confirmed that Nemo-like kinase (NLK) is a tumor suppressor gene. In vitro, it has been found that Nemo-like kinase (NLK) can inhibit cancer by promoting apoptosis of tumor cells. It has been shown that NLK can promote human colon cancer cell DLD-1 cell line. Apoptosis of breast cancer cell line and glioma cell line [1-3]. However, there are few studies of NLK in gliomas, so it can be preliminarily concluded that NLK is a tumor suppressor gene. In addition, in order to find microRNA, that can regulate the expression of NLK, we searched microRNA target gene websites, including TARGETSCAN, MIRANDA, and found that many microRNA, including niR-92b, had potential binding sites on NLK mRNA3'UTR. Therefore, this study focused on the inhibition of glioma proliferation by miR-92b I. Cell cycle progression, the effects of apoptosis and invasion and their possible mechanisms. Methods and results: the study was divided into the following two parts: in the first part, the expression of miR-92b in 5 glioma cell lines and 38 cases of human gliomas of different grades were detected by the method of Real time PCR. The results showed that the expression of miR-92b in glioma tissues and cell lines was higher than that in normal tissues, and there was a positive correlation between the expression of miR-92b and tumor grade. In the second part of the study, miR-92b 鈪，
本文编号：2275836
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