Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum
Exponentially growing, anchorage‐dependent fibroblasts were impeded in their progress through the cell cycle as a result of brief trypsinization from the substratum followed by replating. Untransformed mouse (3T3, clone A31), hamster (CHEF/18‐1) and human (FS2) fibroblasts were partially inhibited f...
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1983
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219541_v114_n1_p53_Campisi http://hdl.handle.net/20.500.12110/paper_00219541_v114_n1_p53_Campisi |
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paper:paper_00219541_v114_n1_p53_Campisi2023-06-08T14:43:46Z Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum radioisotope animal cell autoradiography cell adhesion cell cycle cell cycle s phase cell transformation cytology fibroblast flow cytometry heredity histology human human cell in vitro study mouse Animal Cell Adhesion Cell Cycle Cell Transformation, Neoplastic Cell Transformation, Viral Cytochalasin B Demecolcine DNA DNA Replication Mice Peptide Hydrolases Proteins RNA RNA, Heterogeneous Nuclear Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. Exponentially growing, anchorage‐dependent fibroblasts were impeded in their progress through the cell cycle as a result of brief trypsinization from the substratum followed by replating. Untransformed mouse (3T3, clone A31), hamster (CHEF/18‐1) and human (FS2) fibroblasts were partially inhibited from entering the DNA synthetic (S) phase of the cell cycle for 8 or 12 hours after detachment, even though the cells reattached within an hour of replating and attained a spread morphology 5 or 8 hours later. The decline in the proportion of cells in S phase was accompanied accumulation of cells in G1 as measured by autoradiography and flow microfluorimetry. Cells removed from the substratum by EDTA alone showed identical disturbances of exponential growth. These cell cycle perturbations could be a result of the detachment per se, as opposed to the rounded morphology. Synchronized A31 cells, exposed to colcemid or cytochalasin B for two hours, were not delayed in their entry into S, whereas trypsinization delayed S phase entry by 4 to 5 hours. These drugs disrupt the cytoskeleton without causing detachment. Isotope incorporation experiments revealed no decreases in the rates of protein or RNA synthesis following replating. However, exponentially growing A31 cells, treated for 2 hours with an inhibitor of protein synthesis behaved similarly to those briefly detached from their substratum: 7 hours after treatment, there were fewer cells in S and more cells in G1 relative to untreated cells. Brief treatment with an inhibitor of hn‐RNA synthesis did not alter the cell cycle distribution of these fibroblasts. Three tumorogenic A31 derivatives were less affected by brief detachment from the substratum than were the untransformed cells. The derivative exhibiting the least in vitro growth control (an SV‐40 transformant) showed the least sensitivity to trypsinization, while that derivative having the most in vitro growth control (a Moloney sarcoma virus transformant) was most sensitive. A chemically [benzo(a)pyrene] transformed derivative gave intermediate results with respect to both growth control and sensitivity to detachment from the substratum. The results suggest that as yet unidentified protein(s) necessary for the normal transit through G1 may be quite sensitive to the presence of an anchoring substratum. Copyright © 1983 Wiley‐Liss, Inc. 1983 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219541_v114_n1_p53_Campisi http://hdl.handle.net/20.500.12110/paper_00219541_v114_n1_p53_Campisi |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
radioisotope animal cell autoradiography cell adhesion cell cycle cell cycle s phase cell transformation cytology fibroblast flow cytometry heredity histology human human cell in vitro study mouse Animal Cell Adhesion Cell Cycle Cell Transformation, Neoplastic Cell Transformation, Viral Cytochalasin B Demecolcine DNA DNA Replication Mice Peptide Hydrolases Proteins RNA RNA, Heterogeneous Nuclear Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. |
| spellingShingle |
radioisotope animal cell autoradiography cell adhesion cell cycle cell cycle s phase cell transformation cytology fibroblast flow cytometry heredity histology human human cell in vitro study mouse Animal Cell Adhesion Cell Cycle Cell Transformation, Neoplastic Cell Transformation, Viral Cytochalasin B Demecolcine DNA DNA Replication Mice Peptide Hydrolases Proteins RNA RNA, Heterogeneous Nuclear Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| topic_facet |
radioisotope animal cell autoradiography cell adhesion cell cycle cell cycle s phase cell transformation cytology fibroblast flow cytometry heredity histology human human cell in vitro study mouse Animal Cell Adhesion Cell Cycle Cell Transformation, Neoplastic Cell Transformation, Viral Cytochalasin B Demecolcine DNA DNA Replication Mice Peptide Hydrolases Proteins RNA RNA, Heterogeneous Nuclear Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. |
| description |
Exponentially growing, anchorage‐dependent fibroblasts were impeded in their progress through the cell cycle as a result of brief trypsinization from the substratum followed by replating. Untransformed mouse (3T3, clone A31), hamster (CHEF/18‐1) and human (FS2) fibroblasts were partially inhibited from entering the DNA synthetic (S) phase of the cell cycle for 8 or 12 hours after detachment, even though the cells reattached within an hour of replating and attained a spread morphology 5 or 8 hours later. The decline in the proportion of cells in S phase was accompanied accumulation of cells in G1 as measured by autoradiography and flow microfluorimetry. Cells removed from the substratum by EDTA alone showed identical disturbances of exponential growth. These cell cycle perturbations could be a result of the detachment per se, as opposed to the rounded morphology. Synchronized A31 cells, exposed to colcemid or cytochalasin B for two hours, were not delayed in their entry into S, whereas trypsinization delayed S phase entry by 4 to 5 hours. These drugs disrupt the cytoskeleton without causing detachment. Isotope incorporation experiments revealed no decreases in the rates of protein or RNA synthesis following replating. However, exponentially growing A31 cells, treated for 2 hours with an inhibitor of protein synthesis behaved similarly to those briefly detached from their substratum: 7 hours after treatment, there were fewer cells in S and more cells in G1 relative to untreated cells. Brief treatment with an inhibitor of hn‐RNA synthesis did not alter the cell cycle distribution of these fibroblasts. Three tumorogenic A31 derivatives were less affected by brief detachment from the substratum than were the untransformed cells. The derivative exhibiting the least in vitro growth control (an SV‐40 transformant) showed the least sensitivity to trypsinization, while that derivative having the most in vitro growth control (a Moloney sarcoma virus transformant) was most sensitive. A chemically [benzo(a)pyrene] transformed derivative gave intermediate results with respect to both growth control and sensitivity to detachment from the substratum. The results suggest that as yet unidentified protein(s) necessary for the normal transit through G1 may be quite sensitive to the presence of an anchoring substratum. Copyright © 1983 Wiley‐Liss, Inc. |
| title |
Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| title_short |
Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| title_full |
Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| title_fullStr |
Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| title_full_unstemmed |
Cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| title_sort |
cell cycle perturbations in normal and transformed fibroblasts caused by detachment from the substratum |
| publishDate |
1983 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219541_v114_n1_p53_Campisi http://hdl.handle.net/20.500.12110/paper_00219541_v114_n1_p53_Campisi |
| _version_ |
1768544896211222528 |