MECHANISMS OF EXPRESSION OF DELETED MITOCHONDRIAL DNA FROM CULTURE CELLS

培养细胞中缺失线粒体 DNA 的表达机制

• 批准号: 03670517
• 负责人: KIKUCHI Aiko
• 金额: $ 1.34万
• 依托单位: NATIONAL CENTER OF NEUROLOGY AND PSYCHIATRY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03670517/
• 关键词: MITOCHONDRIA; CULTURED CELLS; DATABASE; CELL BANK; ミトコンドリアサイドパチ-; 培養; 筋細胞; HeLa細胞

Our method developed for cryopreservation and subsequent culture of muscle, skin and lymphocytes could accomplish the maintenance of abnormal mitochondria DNA and the cellular phenotypic characteristics in vitro. The cultured cells including muscle, skin fibro-blast and lymphocytes were more than two thousand samples from two hundred patients suspected with mitochondrail abnormalities by the clinical diagnosis. These cells were used for the analysis of the gene expression of abnormal mitochodrial DNA. We then attempted to develop the software for database about the information of these cells for future use.2. Mutant mitochondrial DNA with large-scale deletions have been frequently observed in patients with chronic progressive external opthalmolplegia(CPEO), a sub group of the mitochondrial encephalomyopathies. To exclude involvement of the nuclear genome in expression of the mitochondrial dysfunction characteristic of CPEO, we introduced the mtDNA of CPEO patient into clonal mtDNA-less HeLa cells, and isolated cybrid clones.3. We attempted to establish cell lines without mitochodria by inactivating mitochondrial DNA with the treatment of ethidium bromide to investigate whether the observed abnormality comes from nuclear genes of mitochondrial genes.

我们开发的肌肉、皮肤和淋巴细胞的冷冻保存和随后的培养方法可以在体外完成异常线粒体DNA和细胞表型特征的维持。培养细胞包括肌肉、皮肤成纤维细胞和淋巴细胞，来自200例临床诊断为线粒体异常的患者的2000多份样本。这些细胞用于分析异常线粒体DNA的基因表达。并尝试开发了细胞信息数据库软件，以备将来使用.慢性进行性眼外肌麻痹（CPEO）是线粒体脑肌病的一个亚组，其线粒体DNA大范围缺失突变是常见的。为了排除CPEO患者线粒体功能异常的核基因组表达，我们将CPEO患者的mtDNA导入克隆的mtDNA缺失的HeLa细胞，分离出胞质杂交体克隆.我们试图建立无线粒体的细胞系，通过用溴化乙锭处理使线粒体DNA失活，以研究观察到的异常是否来自线粒体基因的核基因。

项目成果

NONAKA I.,KOGA Y.,KIKUCHI A., et al.: "Mitochondrial encephalomyopathy and cytochrome c oxydase deficiency:mucle culture study" Acta Neuropathol.82. 286-294 (1991)

NONAKA I.、KOGA Y.、KIKUCHI A. 等人：“线粒体脑肌病和细胞色素 C 氧化酶缺乏症：粘液培养研究”Acta Neuropathol.82。

Haggskhi.J,Ohta.S,Kikuchi,A.et al: "Introduction of disease-relectied mitochondrial DNA deletions into HeLa cells laking mitochondrial DNA results* *in mitochondrial dysfunction" Proc.Natl.Acad.Sci.USA. 88. 10614-10618 (1991)

Haggskhi.J、Ohta.S、Kikuchi,A.等人：“将与疾病相关的线粒体 DNA 缺失引入具有线粒体 DNA 的 HeLa 细胞会导致* *线粒体功能障碍”Proc.Natl.Acad.Sci.USA。

Hayashi J., Ohta, Kikuchi A, et al.: "introduction of disease related mitochondrial DNA deletion into HeLa Cells lacking mitochondrial DNA results in mitochondrial dysfunction" Pro Natl Acad Sci USA. 88. 10614-10618 (1991)

Hayashi J.、Ohta、Kikuchi A 等人：“将与疾病相关的线粒体 DNA 缺失引入缺乏线粒体 DNA 的 HeLa 细胞会导致线粒体功能障碍”Pro Natl Acad Sci USA。

KAMO I.,KUNISHITA T.,KIKUCHI A., et al.: "Mitochondrial encephalomyopathy and cytochrome c oxydase deficiency:mucle culture study" J Immunol.(1993)

KAMO I.、KUNISHITA T.、KIKUCHI A. 等人：“线粒体脑肌病和细胞色素 c 氧化酶缺乏症：粘液培养研究”J 免疫学杂志 (1993)

Kamo I., Kunishita T., Kikuchi A., et al.: "Mitochondrial encephalomyopathy and cytochrome c oxydase deficiency:mucle culture study" J Immunol.(1993)

Kamo I.、Kunishita T.、Kikuchi A. 等人：“线粒体脑肌病和细胞色素 C 氧化酶缺乏症：粘液培养研究”J 免疫学杂志 (1993)