A quantitative system for assay of malignant transformation by chemical carcinogens using a clone derived from BALB/3T3

Kakunaga, Takeo

doi:10.1002/ijc.2910120217

Cited by 287 publications

(116 citation statements)

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“…The transformation frequency obtained with this human cell system is of quite low magnitude compared to the frequencies obtained in many transformation systems using rodent cells, which range from 6 X 10-1 to I X 1O-4 per surviving cell (6,(12)(13)(14)(15).…”

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confidence: 59%

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Neoplastic transformation of human diploid fibroblast cells by chemical carcinogens

Kakunaga

1978

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

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171

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Cultured fibroblast cells derived from a skin biopsy sample taken from normal human adult were exposed to a potent carcinogen, 4-nitroquinoline 1- this observed transformation seemed unlikely because clones of these normal cells could also be used to assess the transforming effect of nitroquinoline oxide. Preliminary results suggest that numerous cell divisions were required for the development of the transformation after nitroquinoline oxide treatment of these human cells.When the transformed cell lines were injected subcutaneously into nude (athymic) mice, solid tumors were produced at the site of inoculation.

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confidence: 59%

“…The cell saturation density was expressed as the cell number per cm2 when counts on 3 successive days did not change significantly. Growth of the suspended cells in soft agar was examined by the procedure described previously (6).…”

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confidence: 99%

Neoplastic transformation of human diploid fibroblast cells by chemical carcinogens

Kakunaga

1978

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

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171

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“…Subclones 3T3-P3 and 3T3-714 were derived from BALB 3T3 A31 cells (14) and described previously (5,15). Both subclones exhibit density-dependent growth, do not grow in soft agar, and are flat with polygonal shapes.…”

Section: Methodsmentioning

confidence: 99%

Mechanisms of Kirsten murine sarcoma virus transformation-induced changes in the collagen phenotype and synthetic rate of BALB 3T3 cells.

Bateman¹,

Peterkofsky²

1981

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

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Specific viral transformation rather than cell selection can explain the previously observed increase in the proportion of type m procollagen compared to type I procollagen in BALB 3T3 cells transformed by Kirsten murine sarcoma virus (Ki-MSV). Two subclones of BALB 3T3 A31 were productively infected with a temperature-sensitive Ki-MSV in the presence of helper murine leukemia virus (MLV), resulting in virtually complete transformation ofcultures and eliminating selection oftransformed foci. Analysis of radioactive collagen, derived from procollagen by pepsin treatment, showed that both of the tsKi-MSV/ MLV-transformed subclones contained a 4-fold greater proportion of type m procollagen than did control MLV-infected cultures. A nonproducer derivative exhibited an even greater change (10-fold), indicating that viral replication was irrelevant. After 48 hr at a nonpermissive temperature, tsKi-MSV-transformed cells retained a high proportion oftype m procollagen, suggesting that either this change is not induced by src protein or else there is a slowly reversible or irreversible step involved. Alternatively, type Im procollagen mRNA may be long lived. In contrast, the relative rate of procollagen synthesis in transformed cells was clearly regulated by src protein. Translation of mRNA from cells preincubated at permissive or nonpermissive temperatures revealed that the decreased relative rate can be explained by a simultaneous small decrease in the level of procollagen mRNA and a large increase in mRNA for noncollagen proteins.Transformation of cells by tumor viruses results in decreased anchorage-and density-dependent growth, changes in cellular morphology (1), and generally a reduction in synthesis of extracellular proteins such as fibronectin (2) and procollagen (3-6). In fibroblasts transformed by Rous sarcoma virus (RSV), these decreases correlate with a reduction in procollagen and fibronectin mRNA (7-9). 3T3 mouse cells transformed by murine sarcoma virus (MSV) also were reported to have a decreased level ofprocollagen mRNA hybridizable to a cDNA probe (10).There are a number ofgenetically distinct collagen types, the properties ofwhich have been reviewed in detail (11,12 The collagen produced by two stable, nontransformed subclones was examined before and after the direct productive infection, and transformation of these cultures with Ki-MSV in the presence of helper murine leukemia virus (MLV), which allows replication of the defective MSV. Comparisons were made with control cultures infected with MLV alone. MSV temperature sensitive for transformation was used in order to determine the role ofthe src gene product. Results demonstrated that MSV transformation, and not merely infection with MLV, induced an increase in the proportion of type III collagen produced. Further experiments were conducted to determine the mechanism of this change. MATERIALS AND METHODSCell Cultures. Subclones 3T3-P3 and 3T3-714 were derived from BALB 3T3 A31 cells (14) and described previously (5, 15). Both subclones exhib...

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“…The standard clonal survival assay used low-density cultures of BALB/ c-3T3 cells and was conducted according to our modification (15) of the method first described by Kakunaga (3). Briefly, 200 WT cells were seeded in either 60-mm culture dishes (Corning Science Products, Corning, NY) or 25-cm2 culture flasks (Corning).…”

Section: Standard Clonal Survival Assaymentioning

confidence: 99%

“…The transformation assay culture vessels were seeded with 3 to assess the reproducibility of dose-related increases of BaP-induced cytotoxic and transforming activities (17). A total of three to six test chemicals were included in each transformation experiment, and each chemical was tested at four treatment doses in two or more independent trials.…”

Section: Transformation Assaymentioning

confidence: 99%

Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

Matthews

Spalding

Tennant

1993

Environ Health Perspect

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This report introduces an improved method of detecting chemical-induced morphological transformation of A-31-1-13 BALB/c-3T3 cells. The new procedure uses an increased target cell population to assess chemicalinduced damage by increasing the initial seeding density and by delaying the initiation time of chemical treatment. Furthermore, a newly developed co-culture clonal survival assay was used to select chemical doses for the transformation assay. This assay measured the relative cloning efficiency (RCE) of chemical treatments in high-density cell cultures. In addition, transformation assay sensitivity was enhanced through the use of improved methods to solubilize many chemicals. From a group of 24 chemicals tested in at least two trials, clear evidence of chemical-induced transformation was detected for 12 chemicals (aphidicolin, barium chloride-2H20, 5-bromo-2'-deoxyuridine, C.I. direct blue 15, trans-cinnamaldehyde, cytosine arabinoside, diphenylnitrosamine, manganese sulfate-H20, 2-mercaptobenzimidazole, mezerein, riddelliine, and 2,6-xylidine); 2 chemicals had equivocal activity [C.I. direct blue 218 and mono(2-ethylhexyl)phthalate], 9 chemicals were inactive [carisoprodol, chloramphenicol sodium succinate, 4-chloro-2-nitroaniline, C.I. acid red 114, isobutyraldehyde, mono(2-ethylhexyl)adipate, sodium fluoride, and 12-O-tetradecanoylphorbol-13-acetate), and 1 chemical had an indeterminate response (2,6-dinitrotoluene). All positive responses were detected in the absence of an exogenous activation system and exhibited significant activity at two or more consecutive doses. This report also presents a mathematical method that uses t-statistics for rank-ordering the potency of chemical-induced transformation responses. This model detects sensitivity differences in experiments used to evaluate chemical-induced transformation. Furthermore, it provides a method to estimate a chemical's transformation response in terms of the historical behavior of the assay, as well as its future activity. The most active of the 24 chemicals was mezerein, and the least active chemical was diphenylnitrosamine.

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A quantitative system for assay of malignant transformation by chemical carcinogens using a clone derived from BALB/3T3

Abstract: A31-714, a subclone isolated from clone A31 of the BALBI3T3 line showed a high degree of contact inhibition and an extremely low incidence of spontaneous transformation, Treatment of this subclone with 4-nitroquinoline-I-oxide, 3-methylcholanthrene, benzo (alpyrene or N-methvl

Cited by 287 publications

References 16 publications

Neoplastic transformation of human diploid fibroblast cells by chemical carcinogens

Neoplastic transformation of human diploid fibroblast cells by chemical carcinogens

Mechanisms of Kirsten murine sarcoma virus transformation-induced changes in the collagen phenotype and synthetic rate of BALB 3T3 cells.

Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

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