Changes in Growth Control and Growth Requirements Associated With Neoplastic Transformation In Vitro12

Paul, Dieter; Henahan, Margaret; Wälter, Stephanie

doi:10.1093/jnci/53.5.1499

Cited by 48 publications

(14 citation statements)

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“…The time between the increase in cGMP and the maximal rate of DNA synthesis after serum addition to quiescent cultures was about 20-24 hr (10), while for a second round of cell division it was only 6-8 hr (Fig. 1), the length of the G1 phase of the 3T3 cell cycle (24). Significantly, cGMP concentrations did not decrease to the original value of resting cultures but were approximately twice as high for about 10 hr or 2 hr, while either resting or growing cells progressed through Go (10) or GI, respectively.…”

Section: Resultsmentioning

confidence: 99%

Cyclic Nucleotides and Growth Control in Cultured Mouse Cells: Correlation of Changes in Intracellular 3′:5′cGMP Concentration with a Specific Phase of the Cell Cycle

Seifert

Rudland

1974

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

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The commencement of cell growth following serum addition to quiescent cultures of mouse fibroblasts is preceded by transient changes in intracellular concentrations of cAMP and cGMP. By artificial depletion of the culture medium for different nutrients, cell growth can be reversibly arrested in various phases of the cell cycle. Here it is shown that the major cGMP increases are only observed when cultures which are arrested in the Go phase are stimulated to grow or when synchronized growing cells pass through the G, phase. In addition to its concomitant decrease, cAMP exhibits rhythmic changes during the cell cycle. This suggests that the increase in cGMP could act as a specific signal for movement of cells out of the Go or G, phase of the cell cycle by activating the pleioltypic and mitogenic program of the cell.Nontransformed fibroblasts in tissue culture usually exist in two reversible growth states, a state of rapid proliferation (growing) and a state of relative quiescence (resting) (t, 2). Reinitiation of growth of quiescent fibroblasts can be induced by addition of fresh serum (3, 4). DNA synthesis is initiated 14-16 hr later in a roughly synchronous manner followed by mitosis and cell division (1). Cells thus reversibly arrested are said to exist in the Go phase of the cell cycle, as the time from mitosis to DNA synthesis for growing cells (GI) is considerably shorter (5) and there are also biochemical differences between cells in Go and G1 (6). The 3':5' cyclic nucleotides cAMP and cGMP were implicated as intracellular signals in the transition from a resting to a growing state, as the cellular concentrations of cAMIP transiently fall (7-10) while those of cGMIP rise after brief exposure of quiescent mouse fibroblasts to animal serum or insulin (10,11,26). Moreover, exogenous additions of high nonphysiological concentrations (10-5-10-3 M) of cAMP or cGMP either prevent the initiation of DNA synthesis by serum (12) or induce substantial increases in DNA synthesis (10) in quiescent cultures, respectively. Growing fibroblasts can also be reversibly arrested by starvation for certain low molecular weight nutrients (13)(14)(15) cGMP, when quiescent mouse fibroblasts, starved for particular nutrients or arrested by serum depletion, are stimulated to reinitiate growth and proceed through the cell cycle. In view of the close analogy between stringent control in bacteria and the pleiotypic response in mammalian cells (16), control of growth by amino-acid levels in the medium is of particular interest. MATERIALS AND METHODSGrowth and Handling of Cells were as described previously (10, 17). Cells were grown at 370 in a 10% CO2 atmosphere in 10 ml of Dulbecco's modified Eagle's medium (DEM) containing 10% calf serum in 9-cm tissue culture dishes unless otherwise stated. Conditions for arrest of cells by nutrient starvation were substantially as reported for 3T3-4A (15) and SV3T3 cells (18). Swiss 3T34A cells plated at 5 X 105 cells per tissue culture dish were grown for 3 days in DEM containing 10% serum. M...

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Section: Resultsmentioning

confidence: 99%

Cyclic Nucleotides and Growth Control in Cultured Mouse Cells: Correlation of Changes in Intracellular 3′:5′cGMP Concentration with a Specific Phase of the Cell Cycle

Seifert

Rudland

1974

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

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“…2Bd). Since the number of cells recovered from these monolayers did not change or decrease after they reached saturation density, this indicates that the transformant was still actively dividing after reaching high cell density, which has often been observed in SV40-transformed fibroblast cells (5,10,25,(28)(29)(30). When REC-A16 was cultured in the presence of a four-hormone mix (Hc + Ss + Tf + ghl) together with added insulin, growth control was regained at higher saturation density (3.0 x 106 cells per 35-mm dish), and clusters of mature adipose cells eventually appeared (Fig.…”

Section: Resultsmentioning

confidence: 77%

“…Although the phenotypic expression of adipocyte differentiation appears normal, REC-A16 cells also exhibited most phenotypes typical of transformed cells when they were cultured in the absence of hormones: (i) they were able to multiply in reduced concentrations of added calf serum, (ii) they acquired the altered property of growth on solid agarose (a weak anchorage dependency for growth), (iii) they grew to higher saturation density than normal cells did, (iv) they did not display growth control at high cell density under usual conditions (loss of contact inhibition), and (v) they expressed an SV40 gene product. These are cellular properties which have most often been observed in SV40-transformed fibroblast-like cells (5,10,28,30).…”

Section: Discussionmentioning

confidence: 97%

“…Simian virus 40 (SV40), a DNA tumor virus, transforms nonpermissive cells and produces cell lines, transformants, which, unlike cells transformed by RNA tumor viruses (10,23,28), are often deficient in their ability to enter the resting state, or stationary phase, at high cell density (5,10,25,29,30). SV40 transformation has been used to study differentiated functions (for a recent review, see reference 50) and to isolate functional cell lines in culture (3, 6-8, 40, 45, 46).…”

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confidence: 99%

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Hormonal regulation of the transformation phenotype in simian virus 40-transformed rat embryonic preadipose cell lines.

Yasumoto¹

1984

Mol. Cell. Biol.

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The regulation of transformed phenotypes was studied in newly isolated preadipose cell lines which were established after infection with simian virus 40 tsA58 dl2009. The clonal cell lines isolated exhibited most of the characteristics typical of transformed cells. The transformants, however, were able to differentiate into adipocytes in the presence of low calf serum (0.5%) and a combination of several hormones, including hydrocortisone and insulin. Treatment with insulin alone stimulated the growth of these cells but did not induce lipid accumulation without added hydrocortisone. The effect of hydrocortisone was accompanied by a restoration of growth control in the transformants after they reached high cell density. The blot hybridization analysis of cellular DNAs digested by restriction enzymes revealed that simian virus 40 genomes were integrated at multiple separate sites at which a head-to-tail oligomeric insertion took place. Large T antigen was synthesized in growing cells but was regulated at high cell density when cells were committed to differentiate by glucocorticoids. These results suggest that the glucocorticoid hydrocortisone is capable of restoring growth regulation at high cell densities to simian virus 40-transformed preadipose cell lines.

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“…This indicates that a serum factor that is depleted by the growth of 3T3 cells is required by BP3T3 cells but is not required by SV3T3 cells. (3,16). In contrast, SV3T3 cells remain distributed throughout the cell cycle, with approximately 50% of the cells in the S phase, even when the growth rate is slowed by a deficiency in serum factors (16).…”

Section: Resultsmentioning

confidence: 99%

Control of growth of benzo(a)pyrene-transformed 3T3 cells.

Holley¹,

Baldwin²,

Kiernan³

et al. 1976

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

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The growth controls observed in benzo [alpyrene-transformed factors (1, 2). The requirement for serum factors is highest during the G1 phase of the cell cycle, prior to the initiation of DNA synthesis (3). Once the cells are committed to initiate DNA synthesis, a much lower serum concentration is required and the lower concentration suffices for the completion of the cell cycle.For 3T3 cells, the serum factors that are needed during G1 to control the initiation of DNA synthesis have been studied extensively (4-7). The serum factors can be replaced, to a large extent, by known materials (4): insulin, a glucocorticoid, and the fibroblast growth factor (FGF) of Gospodarowicz (6).Transformation of 3T3 cells to tumorigenic cells by simian virus 40 (SV40) decreases the serum requirement of the cells (8,9). A low requirement for serum remains for survival of SV3T3 cells (10) and a somewhat higher serum concentration (approximately 0.8%) is required for maximum growth rate.However, requirements for the serum factors that control the initiation of DNA synthesis in 3T3 cells appear to be lost completely in highly transformed SV3T3 cells (8).

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Changes in Growth Control and Growth Requirements Associated With Neoplastic Transformation In Vitro12

Cited by 48 publications

References 0 publications

Cyclic Nucleotides and Growth Control in Cultured Mouse Cells: Correlation of Changes in Intracellular 3′:5′cGMP Concentration with a Specific Phase of the Cell Cycle

Cyclic Nucleotides and Growth Control in Cultured Mouse Cells: Correlation of Changes in Intracellular 3′:5′cGMP Concentration with a Specific Phase of the Cell Cycle

Hormonal regulation of the transformation phenotype in simian virus 40-transformed rat embryonic preadipose cell lines.

Control of growth of benzo(a)pyrene-transformed 3T3 cells.

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