RECOVERY OF THE MISSING TUMORIGENICITY IN MITOCHONDRIAL DNA-LESS HELA-CELLS BY INTRODUCTION OF MITOCHONDRIAL-DNA FROM NORMAL HUMAN-CELLS
RECOVERY OF THE MISSING TUMORIGENICITY IN MITOCHONDRIAL DNA-LESS HELA-CELLS BY INTRODUCTION OF MITOCHONDRIAL-DNA FROM NORMAL HUMAN-CELLS
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DOI：
10.1007/bf01233159
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发表时间：
1992-03-01
期刊：
影响因子：
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通讯作者：
NONAKA, I
中科院分区：
文献类型：
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作者：
HAYASHI, JI;TAKEMITSU, M;NONAKA, I
使用无mtDNA的HeLa细胞检测线粒体DNA （mtDNA）在转化表型表达中的作用。通过Southern blot分析和线粒体合成多肽的[S-35]蛋氨酸标记，证实了在无mtDNA的HeLa细胞系EB8中mtDNA及其产物完全耗尽。裸鼠背部皮下接种1 × 10(7)个EB8细胞，观察EB8细胞的致瘤性。结果表明，HeLa细胞的致瘤性丧失与mtDNA的丢失有很好的对应关系。然而，与HeLa细胞相比，EB8细胞在培养中的生长非常差，这表明EB8细胞明显的致瘤性丧失可能是由于细胞生长不良造成的。将正常人成纤维细胞的mtDNA导入EB8细胞，恢复了EB8细胞缺失的致瘤性和生长。这些观察结果可以解释为mtDNA是致瘤性表达所必需的，但在我们的实验中，mtDNA的突变变化不是表型调节所必需的。
The role of mitochondrial DNA (mtDNA) in the expression of the transformed phenotype was examined using mtDNA-less HeLa cells. Complete depletion of mtDNA and its products in the mtDNA-less HeLa cell line, EB8, was confirmed by Southern blot analysis and by [S-35]methionine labeling of mitochondrially synthesized polypeptides. The tumorigenicity of the EB8 cells was assayed by inoculation of 1 X 10(7) cells subcutaneously into the backs of nude mice. The results showed that the tumorigenicity of HeLa cells was lost in good correspondence with the loss of mtDNA. However, the growth of EB8 cells in culture was very poor compared with that of HeLa cells, indicating that the apparent loss of tumorigenicity in EB8 cells could possibly be due to poor growth of the cells. Introduction of mtDNA from normal human fibroblasts into EB8 cells restored both the missing tumorigenicity and growth of the EB8 cells. These observations could be interpreted to show that mtDNA is required for expression of tumorigenicity, but that mutational changes of the mtDNA are not required for modulation of the phenotype in our experiments.