Transformation and immortalization of human keratinocytes by SV40.
Transformation and immortalization of human keratinocytes by SV40.
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SV40 对人角质形成细胞的转化和永生化。
DOI：
10.1111/1523-1747.ep12540905
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Defendi,V
中科院分区：
文献类型：
--
作者：
Steinberg,ML;Defendi,V
我们已经研究了外观的人表皮角质形成细胞的致癌病毒SV 40感染后的转化属性。感染后不久，只有一小部分细胞对SV 40呈阳性。感染后不久，只有一小部分细胞通过免疫荧光法对SV 40 T抗原呈阳性;这部分细胞的平板接种/集落形成效率和生长速率逐渐增加。细胞化学染色和悬浮于甲基纤维素中诱导的角化细胞包膜形成表明，细胞分化能力逐渐降低。感染细胞进入生长危机期，其特征为细胞病理学和细胞死亡，因为T抗原合成水平在约第10代时达到约90%的阳性细胞。从危机期出现的活细胞被发现表现出锚定非依赖性生长，如在半固体培养基中形成活菌落所示，但认为从锚定非依赖性群体分离的克隆之间的菌落形成存在变异性。出现的人口也表现出表型的不稳定性方面的外观的变体，这与未感染的细胞，表达明确的肌动蛋白细胞骨架。受感染的细胞最终变得“永生化”，如无限期的寿命所证明的，即，复制能力保持远超过未感染细胞的正常衰老时间，我们在体外上皮转化的阶段特异性模型的背景下提出这些发现。
We have studied the appearance of transformed properties following infection of human epidermal keratinocytes by the oncogenic virus SV40. Shortly after infection, only a small fraction of the cells are positive for SV40. Shortly after infections, only a small fraction of the cells are positive for SV40 T antigen by immunofluorescence; this fraction progressively increase in plating/ colony-forming efficiency and growth rate. The capacity of the cells to differentiate progressively decreases, as indicated by cytochemical staining and cornfied cell-envelope formation induced by suspension in methyl cellulose. The infected cells enter a period of growth crisis characterized by cytopathology and cell death as the level of T antigen synthesis reaches about 90 precent positive cells at about the tenth serial passage. Viable cells emerging from the crisis period are found to exhibit anchorage-independent growth, as indicated by the formation of viable colonies in semisolid media, but there is considered variability in colony formation among clones isolated from anchorage-independent populations. The emergent population also manifests phenotypic instability in terms of the appearance of variants, which in contrast to uninfected cells, expresses a well-defined actins cytoskeleton. The infected cells eventually become “immortalized,” as evidenced by an indefinite lifespan, i.e., replication capability maintained well beyond the ordinary time of senescence for uninfected cells, We present these findings in the context of astage-specificmodel of epithelial transformation in vitro.