Montarras D, Poupel O, Fiszman M, Marcovich H
Int J Radiat Oncol Biol Phys. 1985 Feb;11(2):235-9. doi: 10.1016/0360-3016(85)90144-0.
Permanent cell lines of Quail embryo fibroblasts appear in cultures of cells infected with a wild type strain of Rous sarcoma virus (SR-RSV) or with its temperature sensitive transformation mutants (ts-T) (NYts68 and PA101) following a three step process. In step one, infected cells grow twice as fast as the control. The second step consists of a crisis during which the cell population is stationary for four to five weeks. Towards the fourth week several foci of cell growth are observed in the flasks. Respreading of the content of these flasks yields permanent lines. This constitutes the third step of the population evolution. In step one the growth rate of the infected cells is the same irrespective of the incubation temperature (36 degrees C or 41 degrees C) whereas the level of the pp60v-src activity is considerably depressed at 41 degrees C for NYts68 and PA101. Foci do not appear at restrictive temperature in the ts infected population and permanent lines are not recovered under that condition. These lines grow ony at 36 degrees C. It can be shown that the virus which they produce is not modified with respect to the temperature sensitivity of the src gene expression since newly infected fibroblasts grow equally well in step one at both 36 degrees C and 41 degrees C, and stop after the same number of generations. This finding suggests that the events which, during the crisis period, lead to the establishment of permanent lines, take place at the cellular level but depend on the activity of the pp60v-src protein for their occurrence or their expression.
鹌鹑胚胎成纤维细胞的永久细胞系,在感染劳斯肉瘤病毒(SR-RSV)野生型毒株或其温度敏感型转化突变体(ts-T)(NYts68和PA101)的细胞培养物中,经过三个步骤出现。第一步,感染细胞的生长速度是对照细胞的两倍。第二步是危机期,在此期间细胞群体停滞四到五周。到第四周时,在培养瓶中观察到几个细胞生长灶。将这些培养瓶中的细胞内容物重新传代可得到永久细胞系。这构成了群体进化的第三步。在第一步中,无论培养温度(36℃或41℃)如何,感染细胞的生长速度都是相同的,而对于NYts68和PA101,在41℃时pp60v-src活性水平显著降低。在ts感染群体中,在限制温度下不会出现生长灶,在这种条件下也无法获得永久细胞系。这些细胞系仅在36℃生长。可以证明,它们产生的病毒在src基因表达的温度敏感性方面没有改变,因为新感染的成纤维细胞在第一步中在36℃和41℃时生长同样良好,并在相同代数后停止生长。这一发现表明,在危机期导致永久细胞系建立的事件发生在细胞水平,但依赖于pp60v-src蛋白的活性才能发生或表达。