Rat cell line 3y1 and its virogenic polyoma‐ and SV40‐transformed derivatives

Kimura, Genki; Itagaki, Asao; Summers, Jesse

doi:10.1002/ijc.2910150419

Cited by 333 publications

(159 citation statements)

References 34 publications

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“…The rat 3Y1 fibroblastic cell line has been utilized as a model system to study transforming activities of oncogenes (32)(33)(34)(35)(36)(37). Transformed 3Y1 cells bearing adenovirus E1A fragment or simian virus 40 show slow rate of tumorigenesis despite of the increases in both saturation cell density and anchorage-independent growth (33,35).…”

Section: Ha Is a Linear Polysaccharide Composed Of The Repeating Disamentioning

confidence: 99%

Selective Expression and Functional Characteristics of Three Mammalian Hyaluronan Synthases in Oncogenic Malignant Transformation

Itano

Sawai

Atsumi

et al. 2004

Journal of Biological Chemistry

133

102

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Malignant transformation of fibroblasts and epithelial cells is often accompanied by increased hyaluronan production and accumulation. Despite recent progress in the study of hyaluronan biosynthesis, the mechanisms underlying the transformation-induced overproduction of hyaluronan have not been elucidated. Here we report that activity and transcriptional levels of hyaluronan synthase (HAS) significantly increased after oncogenic malignant transformation of a rat 3Y1 fibroblast cell line. Of three HAS isoforms (HAS1, HAS2, and HAS3), only HAS2 gene expression was increased in the v-Ha-ras transformed 3Y1 cells, which show less malignancy. In contrast, both HAS1 and HAS2 expressions were elevated in the highly malignant cells transformed with v-src and/or v-fos. To assess the contribution of HAS expression to the oncogenic malignant transformation, we established stable cell transfectants expressing sense and antisense HAS genes. Antisense suppression of the HAS2 expression significantly decreased hyaluronan production in the cells transformed by the oncogenic v-Ha-ras and eventually led to a reduction in tumorigenicity in the rat peritoneum. The introduction of the HAS1 and HAS2 genes promoted the growth of subcutaneous tumors in a manner dependent on the levels of hyaluronan synthesis. Significant growth promotion was observed within a wide range of HAS1 expression. In contrast, the growth stimulation was only seen within a narrow range of HAS2 expression, and high levels of HAS2 expression even inhibited tumor growth. These results suggest that proper regulation of the expression of each HAS isoform is required for optimal malignant transformation and tumor growth.

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Section: Ha Is a Linear Polysaccharide Composed Of The Repeating Disamentioning

confidence: 99%

Selective Expression and Functional Characteristics of Three Mammalian Hyaluronan Synthases in Oncogenic Malignant Transformation

Itano

Sawai

Atsumi

et al. 2004

Journal of Biological Chemistry

133

102

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“…The results of the CAT assays were quanti®ed by molecular imaging analysis using a Fuji BAS 1000 phosphorimager and luciferase reporter assays were performed in a Lumac Biocounter 1 M2010 luminometer. 3Y1 cells are established Fischer rat embryo ®broblasts (Kimura et al, 1975). Cells were cultured in Dulbecco-modi®ed Eagle's medium (DMEM) supplemented with 10% (v/v) foetal calf serum (FCS), in a humidi®ed atmosphere containing 5% CO 2 at 378C E7 genes on promoters regulating antigen processing and presentation gene expression.…”

mentioning

confidence: 99%

Transcriptional regulation of the major histocompatibility complex (MHC) class I heavy chain, TAP1 and LMP2 genes by the human papillomavirus (HPV) type 6b, 16 and 18 E7 oncoproteins

2000

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We have examined the possibility that the E7 proteins of the high-risk human papillomavirus (HPV) type 16 and 18 and the oncogenic adenovirus (Ad) type 12 E1A protein share the ability to down-regulate the expression of components of the antigen processing and presentation pathway, as a common strategy in the evasion of immune surveillance during the induction of cell transformation. Expression of the HPV 18 E7 oncoprotein, like Ad 12 E1A, resulted in repression of the major histocompatibility complex (MHC) class I heavy chain promoter, as well as repression of a bidirectional promoter that regulates expression of the genes encoding the transporter associated with antigen processing subunit 1 (TAP1) and a proteasome subunit, low molecular weight protein 2 (LMP2). HPV 16 E7 also caused a reduction in class I heavy chain promoter activity, however it did not have any signi®cant e ect on the activity of the bidirectional promoter. Interestingly, expression of the low-risk HPV 6b E7 protein resulted in an increase in MHC class I heavy chain promoter activity, while repressing the TAP1/LMP2 promoter. Interference with the class I pathway could also explain the ability of low-risk HPVs in inducing benign lesions. Oncogene (2000) 19, 4930 ± 4935.

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“…PAb419, a hybridoma cell line producing an antibody against SV40 LT (Harlow et al, 1981), was maintained in ASF104 medium (Ajinomoto) supplemented with 10% fetal calf serum. WI26VA4, CV1, and COS1 were obtained from JCRB cell bank, A640-3Y1 and dl884-3Y1 (Kimura et al, 1975) from RIKEN cell bank with the permission of Dr G Kimura, and PAb419 from RIKEN cell bank with the permission of Dr Y Murakami.…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

p53-independent association between SV40 large T antigen and the major cytosolic heat shock protein, HSP90

Miyata

Yahara

2000

Oncogene

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The simian double strand DNA tumor virus SV40 encodes the 90-kDa multi-functional protein, large T antigen (LT). LT functions by binding to DNA, as well as to many cellular target proteins such as p53 and retinoblastoma protein (pRB). We report here the identi®cation of a cellular heat shock protein, HSP90, as a previously undescribed LT-associated protein.Immunoprecipitates by anti-HSP90 antibodies from LT-expressing cell lysates contained LT protein, as revealed by Western blotting. Conversely, anti-LT antibody co-immunoprecipitated HSP90. Co-immunoprecipitation of HSP90 and LT was observed even after complete immuno-depletion of p53, indicating that the association of LT with HSP90 is p53-independent. LT-HSP90 complexes can be reconstituted from puri®ed HSP90 and unfolded-LT in vitro in an ATP-independent manner but not from HSP90 and native LT, suggesting that non-mature conformation of LT is required for the e cient association with HSP90. Moreover, geldanamycin, an anti-tumor drug that speci®cally binds and inhibits HSP90, reduced the intracellular concentration of LT by destabilizing newly synthesized LT. The above results suggest that HSP90 associates with immature forms of LT both in vivo and in vitro, and thus might assist LT in the formation of a functional, mature structure.

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Rat cell line 3y1 and its virogenic polyoma‐ and SV40‐transformed derivatives

Cited by 333 publications

References 34 publications

Selective Expression and Functional Characteristics of Three Mammalian Hyaluronan Synthases in Oncogenic Malignant Transformation

Selective Expression and Functional Characteristics of Three Mammalian Hyaluronan Synthases in Oncogenic Malignant Transformation

Transcriptional regulation of the major histocompatibility complex (MHC) class I heavy chain, TAP1 and LMP2 genes by the human papillomavirus (HPV) type 6b, 16 and 18 E7 oncoproteins

p53-independent association between SV40 large T antigen and the major cytosolic heat shock protein, HSP90

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