Shiroki K, Segawa K, Koita Y, Shibuya M
Mol Cell Biol. 1986 Dec;6(12):4379-86. doi: 10.1128/mcb.6.12.4379-4386.1986.
Transformed foci were obtained in rat 3Y1 fibroblasts cotransfected with pRmyc 27 (transcriptionally activated c-myc) and pSV2neo DNA. RmycY cell lines (1 to 7) were established from these foci. RmycY cells were small and round and contained enlarged nucleoli in the nucleus. The myc gene was expressed in these cell lines at a much higher level than in 3Y1 cells and at a level similar to that in HL-60 cells. These cell lines formed colonies in soft-agar culture and tumors in syngeneic rats transplanted with RmycY cells. Expression of the gene and colony formation in soft-agar culture were analyzed in subclones from RmycY cell line 1. A correlation between myc gene expression and the ability to form colonies in soft-agar culture was observed in these cells. Antibody against p53 cellular tumor antigen was detected in some sera from tumor-bearing rats. p53 cellular tumor antigen stabilized and accumulated in RmycY cells to the same extent as in simian virus 40-transformed cells. The results suggest that elevated c-myc expression and an increased amount of p53 cause 3Y1 cells to become a more tumorigenic cell line.
将pRmyc 27(转录激活的c-myc)和pSV2neo DNA共转染大鼠3Y1成纤维细胞后获得了转化灶。从这些灶中建立了RmycY细胞系(1至7)。RmycY细胞小而圆,细胞核中含有增大的核仁。myc基因在这些细胞系中的表达水平比在3Y1细胞中高得多,与HL-60细胞中的表达水平相似。这些细胞系在软琼脂培养中形成集落,并在移植了RmycY细胞的同基因大鼠中形成肿瘤。对RmycY细胞系1的亚克隆进行了基因表达和软琼脂培养中集落形成的分析。在这些细胞中观察到myc基因表达与软琼脂培养中形成集落的能力之间存在相关性。在一些荷瘤大鼠的血清中检测到了针对p53细胞肿瘤抗原的抗体。p53细胞肿瘤抗原在RmycY细胞中稳定并积累,程度与猿猴病毒40转化的细胞相同。结果表明,c-myc表达升高和p53量增加导致3Y1细胞成为更具致瘤性的细胞系。