无线粒体DNA的人食管癌细胞系的建立

Establishment of Mitochondrial DNA-Depleted Cell Lines from Human Esophageal Carcinoma Cell

为了研究线粒体疾病呼吸键功能缺陷的分子遗传机制，我们用人食管癌细胞系制备无线粒体DNA的细胞系。在细胞培养液中加溴化乙院50ng／mL、尿嘧啶50μg／mL、丙酮酸100μg／nL，进行连续传代培养。Southern杂交及PCR结果均显示：线粒体DNA进行性减少，呼吸控制率渐降低，到溴化乙啶处理第12天时消失，细胞生长是营养缺陷型，成为无线粒体DNA的细胞系。

In order to study molecular genetic mechanism of deficient respiration capacity of mitochondrialdiseases, we estabIished mitochondrial DNA-depleted cell Iines from human esophageal carcinomacell. The cells were grown in DMEM supplemented with EB (50ng/mL), uridine (50μg/mL),pyruvate (100μg/mL).Results of Southern blots and PCR revealed that mtDNA decreased graduallyand disappeared after EB treatment after 12th days. RCR values were also decreasing and couldn't bedetected in the meantime. Growth of the cells were dependent on uridine and pyruvate which meansthat the cell had become mitochondrial DNA-depleted cell lines.