| 题名 | 新的肺癌相关基因MCRS1 在肿瘤增殖中的探究 |
| 作者 | 周可成 |
| 学位类别 | 硕士 |
| 答辩日期 | 2014-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 曹毅 |
| 关键词 | 非小细胞肺癌 MCRS1 异常高表达 肿瘤细胞增殖 分子机制 |
| 其他题名 | Pathophysiological research of new lung cancer related gene MCRS1, especially in the proliferation |
| 学位专业 | 细胞生物学 |
| 中文摘要 | 肺癌, 常见的恶性肿瘤。随着环境污染和吸烟影响的增加，肺癌的发病率和死亡率迅速上升。目前，肺癌已成为我国恶性肿瘤之首。肿瘤细胞分子水平的改变, 如染色体、基因的改变，已被认为是恶性肿瘤发生的最主要机理。寻找新的肺癌相关基因——癌基因和抑癌基因，对深入研究肺癌发生、发展的病理生理机制，发现新的诊断标记物和治疗靶点有实际意义。 在前期工作中，我们发现微小球核蛋白MCRS1在肺癌组织中表达水平升高尤为明显，统计学分析有非常显著差异（p<0.001）。MCRS1定位于细胞核，参与基因转录调节、细胞转化和恶性肿瘤发生及发展等过程。基于MCRS1在非小细胞肺癌中异常表达，我们首先重点探究其高表达的机制，发现：（1）一定程度上，MCRS1高表达受基因扩增拷贝数变化的影响。（2）MCRS1高表达受miR-129*调控的影响。 我们应用RNA interference技术降低MCRS1的表达，研究其对肿瘤的恶性增殖的影响。（1）通过MTT检测细胞增殖、流式细胞术分析细胞周期分布和Annexin V-FITC/PI双染检测细胞凋亡，其结果显示MCRS1沉默可明显的抑制肺癌细胞增殖、促进细胞凋亡并且延缓细胞周期进程。（2）裸鼠体内成瘤实验显示, MCRS1可以显著影响肿瘤生长能力。这些结果表明，MCRS1在肺癌发生发展过程中发挥重要作用，可能是一个新的肺癌相关基因。 最后，我们通过mRNA基因表达谱芯片、miRNA测序、染色质共沉淀揭示MCRS1促进增殖的机制。我们发现：（1）MCRS1通过影响细胞增殖相关分子的表达促进肺癌的恶性增殖，如CDK、Cyclin、MAPK、Bcl-2、P53、RB等;（2）MCRS1作为重要的转录调控元件，作用于miR-155 host gene 的promoter 区域，调控miRNA-155的表达;（3）通过将ChIP 产物克隆测序的方法，我们得到MCRS1通过活化一些增殖相关基因的promoter直接调控细胞增殖;（4）为了更好地阐述MCRS1的作用机理,我们绘制了miRNA与mRNA的综合调控网络,为将来的后续研究提供一定的参考价值。 关键词：非小细胞肺癌，MCRS1，异常高表达，肿瘤细胞增殖，分子机制 |
| 英文摘要 | Pathophysiological function of oncogenic MCRS1 and its regulation mechanism in non-small cell lung cancer Kecheng Zhou (Cell biology) Directed by Yi Cao Lung cancer is the most common type of cancer in the world. With environment pollution and the increase of smoking, the morbidity and mortality rate of lung cancer were growing rapidly. Nowadays lung cancer has become the leading cause of cancer-related mortality in China. Changes in molecular level, such as chromosomal aberrations, gene mutations and so on, have been recognized as the most significant mechanisms in the genesis and development of cancer. Searching for new tumor suppressor genes or oncogenes do really benefits to the research of lung cancer and its mechanisms, with practical significance indeed. In our previous work, overexpression of MCRS1 was confirmed in lung cancer tissues and cell lines. MCRS1, microspherule protein 1, localized to the nucleus with known roles in transcription regulation, cellular transformation, and tumorigenesis. Since the abnormal high expression of MCRS1 in lung cancer, we focus on the reasons firstly. The results show: (1) MCRS1 amplification was mediated by the alteration in DNA copy number at an certain degree. (2) miR-129* down-regulation in NSCLCs can affect the high expression of MCRS1. RNA interfere were adopted to decrease the expression of MCRS1, and further analyses its promotion in proliferation and tumorigenesis. (1) The data revealed that MCRS1 silencing inhibited cell proliferation, increased apoptosis, and induced cell cycle arrest at the G1 phase in lung cancer cells. (2) The data in vivo inform us that MCRS1 can greatly affect the tumor growth in nude mice. All the results give evidence that MCRS1 has a significant role in cancer development, indicating it could be a newly found lung cancer related gene. In addition, to further explore signal transduction pathways associated with MCRS1 activity in NSCLCs，we performed mRNA , microRNA (miRNA) expression profiles and Chromatin Immunoprecipitation (ChIP assay). We discovered: (1) Downstream effectors were clarified by analysis of mRNA profiles and confirmed by the q-RT-PCR , such as CDK, Cyclin、MAPK、Bcl-2、P53、RB and so on; (2) As an vital transcriptional factor, MCRS1 can bind to the promoter region of miR-155 host gene (MIR155HG); (3) After ChIP assay, gene cloning and sequencing, we got some genes that can be activated by MCRS1, which have essential roles in proliferation; (4) To better understand the regulation web and functions of MCRS1, we mapped the interactive regulation web by bioinformatics analysis of the finding of changed mRNA and miRNA, which provided reference for further related research. Keywords: Non-small cell lung cancer (NSCLC), MCRS1, Abnormal Overexpression, Cell proliferation, Molecular mechanism. |
| 语种 | 中文 |
| 公开日期 | 2014-07-01 |
| 内容类型 | 学位论文 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7923]  |
| 专题 | 昆明动物研究所_分子病理学 |
| 推荐引用方式 GB/T 7714 | 周可成. 新的肺癌相关基因MCRS1 在肿瘤增殖中的探究[D]. 北京. 中国科学院研究生院. 2014. |
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