| Massive programmed cell death in intestinal epithelial cells induced by three-dimensional growth conditions: suppression by mutant c-H-ras oncogene expression. | |
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MedLine Citation:
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PMID: 8522614 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Deregulation of molecular pathways controlling cell survival and death, including programmed cell death, are thought to be important factors in tumor formation, disease progression, and response to therapy. Studies devoted to analyzing the role of programmed cell death in cancer have been carried out primarily using conventional monolayer cell culture systems. However the majority of cancers grow as three-dimensional solid tumors. Because gene expression, and possibly function, can be significantly altered under such conditions, we decided to analyze the control and characteristics of cell death using a compatible three-dimensional tissue culture system (multicellular spheroids) and compare the results obtained to those using two-dimensional monolayer cell culture. To do so we selected for study an immortalized, but nontumorigenic line of rat intestinal epithelial cells, called IEC-18, and several tumorigenic variants of IEC-18 obtained by transfection with a mutant (activated) c-H-ras oncogene. The rationale for choosing these cell lines was based in part on the fact that intestinal epithelial cells grow in vivo in a monolayer-like manner and form solid tumors only after sustaining certain genetic mutations, including those involving the ras gene family. We found that the IEC-18 cells, which grow readily and survive in monolayer cell culture, undergo massive cell death within 48-72 h when cultured as multicellular spheroids on a nonadhesive surface. This process was accompanied by a number of features associated with programmed cell death including chromatin condensation (Hoechst 33258 staining) apoptotic morphology, DNA degradation, and a virtual complete loss of colony forming (clonogenic) ability in the absence of apparent membrane damage as well as accumulation of lipid containing vacuoles in the cytoplasm. Moreover, enforced over-expression of a transfected bcl-2 gene could prevent this cell death process from taking place. In marked contrast, three different stably transfected ras clones of IEC-18 survived when grown as multicellular spheroids. In addition, an IEC cell line (called clone 25) carrying its mutant transfected ras under a glucocorticoid inducible promoter survived in three-dimensional culture only when the cells were exposed to dexamethasone. If exposure to dexamethasone was delayed for as long as 48 h the cells nevertheless survived, whereas the cells became irreversibly committed to programmed cell death (PCD) if exposed to dexamethasone after 72 h. These results suggest that intestinal epithelial cells may be programmed to activate a PCD pathway upon detachment from a physiologic two-dimensional monolayer configuration, and that this process of adhesion regulated programmed cell death (ARPCD) can be substantially suppressed by expression of a mutant ras oncogene.(ABSTRACT TRUNCATED AT 400 WORDS) |
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Authors:
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J Rak; Y Mitsuhashi; V Erdos; S N Huang; J Filmus; R S Kerbel |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: The Journal of cell biology Volume: 131 ISSN: 0021-9525 ISO Abbreviation: J. Cell Biol. Publication Date: 1995 Dec |
Date Detail:
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Created Date: 1996-01-23 Completed Date: 1996-01-23 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 0375356 Medline TA: J Cell Biol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 1587-98 Citation Subset: IM |
Affiliation:
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Division of Cancer Biology Research, Reichmann Research Building, Sunnybrook Health Science Centre, Toronto, Ontario. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Apoptosis / physiology* Cell Adhesion / physiology Epithelial Cells Epithelium / physiology Gene Expression Regulation / physiology Intestines / cytology* Mutation / physiology Protein-Tyrosine Kinases / physiology Proto-Oncogene Proteins / physiology Proto-Oncogene Proteins c-bcl-2 Proto-Oncogene Proteins p21(ras) / genetics* Rats Spheroids, Cellular / cytology Time Factors Tumor Cells, Cultured / cytology |
| Grant Support | |
ID/Acronym/Agency:
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CA-41223/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Proto-Oncogene Proteins; 0/Proto-Oncogene Proteins c-bcl-2; EC 2.7.10.1/Protein-Tyrosine Kinases; EC 3.6.5.2/Proto-Oncogene Proteins p21(ras) |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
| Full Text | |
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Journal Information Journal ID (nlm-ta): J Cell Biol ISSN: 0021-9525 ISSN: 1540-8140 Publisher: The Rockefeller University Press |
Article Information Download PDF  Print publication date: Day: 2 Month: 12 Year: 1995 Volume: 131 Issue: 6 First Page: 1587 Last Page: 1598 ID: 2120690 Publisher Id: 96101936 PubMed Id: 8522614 |
| Massive programmed cell death in intestinal epithelial cells induced by three-dimensional growth conditions: suppression by mutant c-H-ras oncogene expression | |
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