miR-497抑制脑胶质瘤细胞增殖的作用研究
发布时间：2018-11-16 13:41
【摘要】：近年来的研究表明SALL4在许多种肿瘤中高表达，是候选的原癌基因。然而有关miRNA与SALL4的调控关系的研究报道较少，SALL4在肿瘤中的表达调控机制还不是很清楚。我们用qRT-PCR技术分析了miR-497在50例脑胶质瘤组织样品中的表达情况，结果表明84%（42/50）的肿瘤组织样品中miR-497的表达水平显著低于对照组，结合课题组前期SALL4在同种胶质瘤中的表达水平实验结果，并通过miR-497与SALL4的表达相关性分析显示，miR-497与SALL4具有显著的负相关性。本论文通过生物信息预测发现SALL4是miR-497的靶基因。应用双荧光素酶报告系统、qRT-PCR和western-blot技术发现，miR-497的过表达在mRNA水平和蛋白水平均显著抑制SALL4的表达，确认SALL4为miR-497的靶基因。通过MTT增殖曲线、软琼脂集落形成实验，我们发现miR-497对于脑胶质瘤细胞的增殖能力具有显著抑制作用。为了探讨miR-497抑制胶质瘤增殖的作用机制，，我们通过PI-Annexin V双染及Caspase-3/7活性分析显示，miR-497可以诱导脑胶质瘤细胞的凋亡。同时应用western blot技术检测FADD的表达，结果显示miR-497过表达的脑胶质瘤细胞中，FADD显著高表达。 上述实验结果表明，miR-497可以通过调控SALL4的表达来诱导细胞凋亡并抑制脑胶质瘤细胞的增殖能力。
[Abstract]:Recent studies have shown that SALL4 is highly expressed in many kinds of tumors and is a candidate proto-oncogene. However, there are few studies on the regulatory relationship between miRNA and SALL4, and the mechanism of SALL4 expression in tumor is not clear. We analyzed the expression of miR-497 in 50 cases of glioma by qRT-PCR technique. The results showed that the expression of miR-497 in 84% (42 / 50) of tumor tissues was significantly lower than that in the control group. According to the experimental results of the expression of SALL4 in the same glioma and the correlation analysis of the expression of miR-497 and SALL4, it was found that there was a significant negative correlation between miR-497 and SALL4. In this paper, we found that SALL4 is the target gene of miR-497 by biological information prediction. Using double luciferase reporting system, qRT-PCR and western-blot techniques, it was found that the overexpression of miR-497 significantly inhibited the expression of SALL4 at both mRNA and protein levels. SALL4 was identified as the target gene of miR-497. Through the MTT proliferation curve and soft Agar colony formation test, we found that miR-497 has a significant inhibitory effect on the proliferation of glioma cells. In order to investigate the mechanism of miR-497 inhibiting glioma proliferation, PI-Annexin V double staining and Caspase-3/7 activity analysis showed that miR-497 could induce apoptosis of glioma cells. At the same time, the expression of FADD was detected by western blot technique. The results showed that FADD was significantly overexpressed in glioma cells with overexpression of miR-497. These results suggest that miR-497 can induce apoptosis and inhibit the proliferation of glioma cells by regulating the expression of SALL4.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41
本文编号：2335658
[Abstract]:Recent studies have shown that SALL4 is highly expressed in many kinds of tumors and is a candidate proto-oncogene. However, there are few studies on the regulatory relationship between miRNA and SALL4, and the mechanism of SALL4 expression in tumor is not clear. We analyzed the expression of miR-497 in 50 cases of glioma by qRT-PCR technique. The results showed that the expression of miR-497 in 84% (42 / 50) of tumor tissues was significantly lower than that in the control group. According to the experimental results of the expression of SALL4 in the same glioma and the correlation analysis of the expression of miR-497 and SALL4, it was found that there was a significant negative correlation between miR-497 and SALL4. In this paper, we found that SALL4 is the target gene of miR-497 by biological information prediction. Using double luciferase reporting system, qRT-PCR and western-blot techniques, it was found that the overexpression of miR-497 significantly inhibited the expression of SALL4 at both mRNA and protein levels. SALL4 was identified as the target gene of miR-497. Through the MTT proliferation curve and soft Agar colony formation test, we found that miR-497 has a significant inhibitory effect on the proliferation of glioma cells. In order to investigate the mechanism of miR-497 inhibiting glioma proliferation, PI-Annexin V double staining and Caspase-3/7 activity analysis showed that miR-497 could induce apoptosis of glioma cells. At the same time, the expression of FADD was detected by western blot technique. The results showed that FADD was significantly overexpressed in glioma cells with overexpression of miR-497. These results suggest that miR-497 can induce apoptosis and inhibit the proliferation of glioma cells by regulating the expression of SALL4.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41
【参考文献】
相关期刊论文 前1条
1 陈忠平,周旺宁;我国胶质瘤诊断治疗现状和努力方向[J];中国肿瘤;2005年02期
本文编号：2335658
本文链接：https://www.wllwen.com/yixuelunwen/shenjingyixue/2335658.html
最近更新
教材专著
