Oda K, Masuda-Murata M, Shiroki K, Handa H
J Virol. 1986 Apr;58(1):125-33. doi: 10.1128/JVI.58.1.125-133.1986.
Several lines of rat 3Y1 cells in which expression of the adenovirus type 12 E1A gene can be regulated by dexamethasone were established by introduction of recombinant vector DNA containing the adenovirus type 12 E1A gene placed downstream of the hormone-inducible promoter of mouse mammary tumor virus. These cell lines (gMA cells) produced low basal levels of the E1A transcripts and proteins in normal medium and much higher levels upon addition of dexamethasone to the medium. When dexamethasone was added to density-arrested cells, DNA synthesis was induced in 10 to 40% of the cells, the percentage depending on the cell line. DNA synthesis was increased to up to 60% of the cell population by further addition of epidermal growth factor. Indirect immunofluorescence detection of E1A proteins in gMA cells treated with dexamethasone indicated that the intensity of fluorescence in cells varied and that the proportion of cells synthesizing DNA was correlated with the proportion that exhibited strong fluorescence. These results indicate that the E1A gene has a function to trigger the synthesis of cellular DNA.
通过导入含有置于小鼠乳腺肿瘤病毒激素诱导型启动子下游的腺病毒12型E1A基因的重组载体DNA,建立了几株大鼠3Y1细胞系,其中腺病毒12型E1A基因的表达可受地塞米松调控。这些细胞系(gMA细胞)在正常培养基中产生低水平的E1A转录本和蛋白质,而在培养基中添加地塞米松后水平会高得多。当地塞米松添加到密度停滞的细胞中时,10%至40%的细胞会诱导DNA合成,具体百分比取决于细胞系。通过进一步添加表皮生长因子,DNA合成增加到细胞群体的60%。用地塞米松处理的gMA细胞中E1A蛋白的间接免疫荧光检测表明,细胞中的荧光强度各不相同,并且合成DNA的细胞比例与显示强荧光的细胞比例相关。这些结果表明,E1A基因具有触发细胞DNA合成的功能。