Human cell transformation by simian virus 40—A review

Sack, George H.

doi:10.1007/bf02618025

In Vitro

1981

DOI: 10.1007/bf02618025

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Human cell transformation by simian virus 40—A review

George H. Sack

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1986

2001

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Cited by 127 publications

(62 citation statements)

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“…Constitutive expression of the DNA tumor virus SV40 T-antigen is the most effective means to immortalize primary human cells, or at least to prolong their life span. [25][26][27] One limitation is that SV40 T-antigen transformation preferentially hits actively proliferating cells, or at least cells that still have the capacity to enter the cell cycle. T antigen can interact with cellular proteins controlling progression through the cell cycle such as the pocket proteins Rb-1, p107, and p130 28 -30 as well as p53.…”

mentioning

confidence: 99%

Identification of an Invasive, N-Cadherin-Expressing Epithelial Cell Type in Endometriosis Using a New Cell Culture Model

Zeitvogel

Baumann

Starzinski‐Powitz

2001

The American Journal of Pathology

244

226

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mentioning

confidence: 99%

Identification of an Invasive, N-Cadherin-Expressing Epithelial Cell Type in Endometriosis Using a New Cell Culture Model

Zeitvogel

Baumann

Starzinski‐Powitz

2001

The American Journal of Pathology

244

226

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“…Phenotypic characteristics have been well maintained in SV40-transformed fibroblasts (13)(14)(15). In SV40-transformed epithelial cells (16)(17)(18)(19), certain aspects of phenotype are retained better than others (17,18,20,21).…”

mentioning

confidence: 99%

Characterization of human tracheal epithelial cells transformed by an origin-defective simian virus 40.

Gruenert

Basbaum

Welsh

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

211

148

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To facilitate understanding of the mechanisms underlying pulmonary diseases, including lung cancer and cystic fibrosis, we have transformed and characterized cultures of human tracheal epithelial cells. Cells were transfected by calcium phosphate precipitation with a plasmid containing a replication-defective simian virus 40 (SV40) genome. Colonies of cells with enhanced growth potential were isolated and analyzed for transformation-and epithelialspecific characteristics. Precrisis cells were observed to express the SV40 large tumor antigen, produce cytokeratins, have microvilli, and form tight junctions. After crisis, cells continued to express the SV40 large tumor antigen as well as epithelial-specific cytokeratins and to display the apical membrane microvilli. Apical membrane Cl channels were opened in postcrisis cells exposed to 50 ,LM forskolin. These channels showed electrical properties similar to those observed in primary cultures. The postcrisis cells have been in culture for >250 generations and are potentially "immortal." In addition to providing a useful in vitro model for the study of ion transport by human airway epithelial cells, the cells can be used to examine stages of neoplastic progression. (6,7).In contrast to the relatively large number of studies describing cells from other organs (recently reviewed in refs. 1 and 2), only three reports indicate transformation of respiratory tract cells (8-10). Bronchial epithelial cells immortalized with v-Ha-ras display anchorage-independent growth and form tumors in nude mice. However, these cells were poorly differentiated and did not contain tight junctions or keratins characteristic of differentiated epithelial cells. The epithelial origin of the cells was verified by the presence of keratin. A somewhat greater level of differentiation was retained by diethylnitrosamine-transformed fetal tracheal cells, which contained mucin. These cells also expressed carcinoembryonic antigen and wool merokeratin and had enhanced, although limited, growth capacity (9). A third study primarily described methods for transfection and showed expression of the SV40 large tumor antigen (T antigen) three to four subcultures posttransfection (10).A goal of the present study was to develop a transformed human tracheal epithelial (HTE) cell line for detailed studies of the mechanisms regulating Cl ion transport. This aspect of epithelial function is essential for rehydration of airway mucus. Abnormalities in the Cl conductance pathway also appear critical to disease processes such as those occurring in cystic fibrosis (11,12).Phenotypic characteristics have been well maintained in SV40-transformed fibroblasts (13-15). In SV40-transformed epithelial cells (16)(17)(18)(19), certain aspects of phenotype are retained better than others (17,18,20,21). A plasmid containing a SV40 genome defective in the origin of replication (pSVori-) has been shown to enhance the transformation efficiency of human fibroblasts (22). The defective origin of replication precludes background ...

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“…Simian virus 40 (SV40) is a small DNA virus able to infect HDF resulting in both transformation and virus production (39,42). Initially, a small number of infected cells produce SV40 T antigen, but with passage the percentage of T-positive cells increases and acquires the phenotypic characteristic of transformation (10,22).…”

mentioning

confidence: 99%

“…The SV40 HDF system offers a unique opportunity to investigate the development of the neoplastic phenotype of a human diploid cell with limited life span which can be modulated by the presence andor integration of the SV40 genome (39). The evidence indicates that the SV40 early gene product, large T, is most important in the initiation and establishment of cell lines with a tumorigenic phenotype.…”

mentioning

confidence: 99%

Correlation of DNA content, p53, T antigen, and V antigen in simian virus 40‐infected human diploid cells

1989

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Human diploid fibroblasts (HDF) have a finite life span in cell culture which can be extended when transformed with simian virus 40 (SV40). Flow cytometric analysis of SV40-HDF transformation allowed DNA content changes to be correlated with the appearance, quantity, and distribution of T antigen, p53, and V antigen, three proteins associated with this process. These studies demonstrated a shift in the DNA content to tetraploidy, which was correlated with the age of the SV4O-HDF but not the time of infection. A significant increase of the epitope recognized by PAbl22 to host p53 and the epitope PAblOl to SV40 T antigen occurred at the same time the tetraploid population appeared. However, an antigen reactive with SV40 V antibody was present at high levels in most of the population early after infection, but the levels declined with time. The percentage of PAblOl-T antigen-positive cells increased more rapidly in cells infected at a late passage, and this was concomitant with the shift in DNA content to tetraploid. Analysis of the mean fluorescence of total, gated populations (GI, G2, and > Gt) demonstrated that a threshold level of p53 and T antigen was reached in each compartment of the cell cycle. As the transformed phenotype appeared, a population of cells was continually released into the supernatant, and although these cells had a DNA pattern similar to the monolayer cells, the T antigen and p53 levels were 3-5 times higher in the tetraploid 6 2 cells.These studies correlated the expression of proteins associated with viral transformation in HDF which vary with time and shift in DNA content.

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Human cell transformation by simian virus 40—A review

Cited by 127 publications

References 173 publications

Identification of an Invasive, N-Cadherin-Expressing Epithelial Cell Type in Endometriosis Using a New Cell Culture Model

Identification of an Invasive, N-Cadherin-Expressing Epithelial Cell Type in Endometriosis Using a New Cell Culture Model

Characterization of human tracheal epithelial cells transformed by an origin-defective simian virus 40.

Correlation of DNA content, p53, T antigen, and V antigen in simian virus 40‐infected human diploid cells

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