Raj G, Gordon J, Logan T, Hall D, Deluca A, Giordano A, Khalili K
THOMAS JEFFERSON UNIV,JEFFERSON INST MOLEC MED,DEPT BIOCHEM & MOLEC BIOL,MOLEC NEUROVIROL SECT,PHILADELPHIA,PA 19107. THOMAS JEFFERSON UNIV,JEFFERSON CANC INST,DEPT MICROBIOL & IMMUNOL,PHILADELPHIA,PA 19107.
Int J Oncol. 1995 Oct;7(4):801-8. doi: 10.3892/ijo.7.4.801.
Intracerebral injection of human polyomavirus, JCV, into neonatal hamsters causes tumors of,glial origin. HJC is an established cell Line derived from a JCV-induced mixed hamster brain tumor with astrocytic and ependymal components. Flow cytometric and immunohistochemical analysis of HJC suggests that it is comprised of a mixed population of cells all of which contain the JCV early protein, T-antigen, in the nuclei. Five individual clonal lines, called HJC-15a to HJC-15e, were isolated by limiting dilution and were found to exhibit distinct morphological characteristics with 25-30% variation in their sizes. It was evident that each clone has unique growth rates, doubling times, and cell cycle parameters with different G(1), S, and G(2) phase times. All clonal cells showed the presence of the JCV early protein in the nucleus. Of interest was the observation from immunoprecipitation and Western analysis indicating qualitative and quantitative differences in the T-antigen isoforms produced in these cells. Similar to the parental clone, HJC-15b produced two distinct forms of JCV T-antigen isoforms, 88 kDa and 92 kDa proteins. In addition, HJC-15c was able to produce a 23-25 kDa protein which was recognized by anti-T-antigen antibody. The activity of cyclin-dependent kinases, in particular cdc2, was higher in HJC-15c than in the other cell lines. The data presented herein indicates that glioblastomas induced by viral T-antigen expression are composed of a multitude of distinct cells that possess a variety of different characteristics.
将人多瘤病毒JCV脑内注射到新生仓鼠体内会引发神经胶质起源的肿瘤。HJC是一个已建立的细胞系,源自JCV诱导的具有星形细胞和室管膜成分的混合仓鼠脑肿瘤。对HJC进行的流式细胞术和免疫组织化学分析表明,它由一群混合细胞组成,所有细胞的细胞核中都含有JCV早期蛋白T抗原。通过有限稀释法分离出五个单独的克隆系,分别称为HJC - 15a至HJC - 15e,发现它们具有明显的形态特征,大小差异为25% - 30%。很明显,每个克隆都有独特的生长速率、倍增时间和细胞周期参数,其G1、S和G2期时间不同。所有克隆细胞的细胞核中都显示存在JCV早期蛋白。有趣的是,免疫沉淀和Western分析观察结果表明,这些细胞产生的T抗原异构体在定性和定量上存在差异。与亲本克隆相似,HJC - 15b产生两种不同形式的JCV T抗原异构体,即88 kDa和92 kDa蛋白。此外,HJC - 15c能够产生一种23 - 25 kDa的蛋白,该蛋白可被抗T抗原抗体识别。HJC - 15c中细胞周期蛋白依赖性激酶,特别是cdc2的活性高于其他细胞系。本文提供的数据表明,由病毒T抗原表达诱导的胶质母细胞瘤由众多具有各种不同特征的独特细胞组成。