Role of inactivation of components of mitochondrial complex II in the formation of paragangliomas/pheochromcytomas

线粒体复合物 II 成分失活在副神经节瘤/嗜铬细胞瘤形成中的作用

• 批准号: 5402375
• 负责人: Professor Dr. Ulrich Müller
• 金额: --
• 依托单位: Institut für Humangenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2005-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5402375?language=en
• 关键词: Role; inactivation; components; mitochondrial; complex

Mutations in genes coding for subunits B,C, and D of mitochondrial complex II (SDHB, SDHC, SDHD) can cause paragangliomas and pheochromocytomas. SDHB, SDHC, and SDHD appear to function as tumor suppressor genes. A germ line mutation plus a deletion of the wild-type allele in target tissues (neural-crest-derived tissues) result in tumor formation. This mechanism was clearly demonstrated for SDHC and SDHD, and somewhat less convincingly für SDHB mutations. This application aims at the elucidation of molecular mechanisms resulting in tumor formation in the absence of either SDHB, C, or D. The respective genes will be inactivated in both PC12 and NIH/3T3 cells by small interfering RNAs (siRNA) using a plasmid-based system. The cells will be investigated for colony formation by soft agar assays and the ultrastructural morphology, in paricular that of the mitochondria, will be studied by electron microscopy. Respiratory chain enzymes will be tested to investigate wheter the knock-downs resulted in loss of complex II enzymatic activity. Expression pattern of genes will be investigated by microarray and realtime quantitative PCR analyses. Special emphasis will be placed on genes regulated by hypoxia inducible factor 1a and those involved in the cell cycle.

线粒体复合体II （SDHB， SDHC， SDHD）编码亚基B，C和D的基因突变可导致副神经节瘤和嗜铬细胞瘤。SDHB、SDHC和SDHD似乎具有肿瘤抑制基因的功能。在靶组织（神经冠源性组织）中，种系突变加上野生型等位基因的缺失导致肿瘤的形成。这种机制在SDHC和SDHD中得到了明确的证明，而在r SDHB突变中则不那么令人信服。本应用程序旨在阐明在缺乏SDHB、C或d的情况下导致肿瘤形成的分子机制。使用基于质粒的系统，通过小干扰rna （siRNA）在PC12和NIH/3T3细胞中使各自的基因失活。这些细胞将通过软琼脂实验研究集落形成，并通过电子显微镜研究超微结构形态，特别是线粒体的超微结构形态。将对呼吸链酶进行测试，以研究这种敲除是否导致复合物II酶活性的丧失。基因的表达模式将通过微阵列和实时定量PCR分析进行研究。特别强调将放在基因调控的缺氧诱导因子1a和那些参与细胞周期。