Akagi Tsuyoshi, Sasai Ken, Hanafusa Hidesaburo
Laboratory of Molecular Oncology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka 565-0874, Japan.
Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13567-72. doi: 10.1073/pnas.1834876100. Epub 2003 Nov 3.
Human cells are known to be more refractory than rodent cells against oncogenic transformation in vitro. To date, the molecular mechanisms underlying such resistance remain largely unknown. The combination of simian virus 40 early region and H-Ras V12 has been effective for transformation of rat embryo fibroblasts, but not for human cells. However, the additional ectopic expression of the telomerase catalytic subunit (hTERT) was reported to be capable of causing transformation of normal human cells. In this study, however, we demonstrate that the combined expression of the above-mentioned three genetic elements is not always sufficient to transform normal human diploid fibroblasts (HDF). Although the expression and function of these introduced genetic elements were essentially the same, among four HDF, TIG-1 and TIG-3 were resistant to transformation. The other two (BJ and IMR-90) showed transformed phenotypes, but they were much restricted compared with rat embryo fibroblasts in expressing simian virus 40 early region and H-Ras V12. In correlation with these phenotypes, TIG-1 and TIG-3 remained diploid after the introduction of these genetic elements, whereas BJ and IMR-90 became highly aneuploid. These results strongly suggest that the lack of telomerase is not the sole reason for the refractory nature of HDF against transformation and that normal human cells have still undefined intrinsic mechanisms rendering them resistant to oncogenic transformation.
已知在体外,人类细胞比啮齿动物细胞对致癌转化更具抗性。迄今为止,这种抗性背后的分子机制仍 largely 未知。猿猴病毒40早期区域和H-Ras V12的组合对大鼠胚胎成纤维细胞的转化有效,但对人类细胞无效。然而,据报道端粒酶催化亚基(hTERT)的额外异位表达能够导致正常人细胞的转化。然而,在本研究中,我们证明上述三种遗传元件的联合表达并不总是足以转化正常人二倍体成纤维细胞(HDF)。尽管这些导入的遗传元件的表达和功能基本相同,但在四个HDF中,TIG-1和TIG-3对转化具有抗性。另外两个(BJ和IMR-90)表现出转化表型,但与大鼠胚胎成纤维细胞相比,它们在表达猿猴病毒40早期区域和H-Ras V12方面受到很大限制。与这些表型相关,TIG-1和TIG-3在导入这些遗传元件后仍保持二倍体状态,而BJ和IMR-90则变为高度非整倍体。这些结果强烈表明,端粒酶的缺乏不是HDF对转化具有抗性的唯一原因,并且正常人细胞仍然具有未定义的内在机制,使其对致癌转化具有抗性。