Selective Activation of Ha-rasval12Oncogene Increases Susceptibility of NIH/3T3 Cells to TNF-α
Abstract
This is the first report demonstrating that NIH/3T3 fibroblasts utilize the Raf-1/MAPK pathway to sensitize themselves to tumor necrosis factor-alpha (TNF-alpha) cytotoxicity under Ha-rasVal12 oncogene-overexpressed conditions. This paper clearly shows that the sensitivity of NIH/3T3 cells to TNF-alpha cytotoxicity positively correlated with the expression level of activated Ha-ras transgene, which was manipulated either positively by isopropyl-beta-d-thiogalactoside (IPTG) induction or negatively by a ribozyme or a dominant negative Ras suppression. Further analysis revealed that after TNF-alpha treatment, Ha-ras-overexpressed transformants underwent apoptosis. Overexpression of dominant negative Raf-1, Rac1, or RhoA in the Ha-ras transformants clarified that among these factors, only dominant negative Raf-1 could reverse the cell sensitivity to TNF-alpha, indicating that Raf-1, as a proapoptotic factor, indeed participates in TNF-alpha cytotoxicity. The anti-apoptotic roles of Bcl-2 and PI(3) kinase are also demonstrated by the Ha-ras transformants which became more resistant to TNF-alpha while overexpressing Bcl-2 or the activated p110 catalytic subunit. The analyses of the cell cycle and nuclear transcription factor activities revealed that TNF-alpha treatment caused the Ha-ras overexpressed transformants to shift from S to G0/G1 phase and increased the responses of AP-1, c-fos, and c-myc. Taken together, we suggest that the possible action of Ha-ras overexpression to sen…
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NIH3T3 cells are highly responsive to TNFα and express endogenous Mfn2.
“In one set of studies we selected NIH3T3 cells, since they are highly responsive to TNF␣ (42) and express endogenous Mfn2 (Fig. 7).”
From Expression of <i>Mfn2</i>, the Charcot-Marie-Tooth Neuropathy Type 2A Gene, in Human Skeletal Muscle (2005) · cites this paper as [42]
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- Authors
- Chang, M.-Y., Won, S.-J., Yang, B.-C., Jan, M.-S., & Liu, H.-S.
- Affiliations
- National Cheng Kung University (TW)
- Corresponding author
- Liu, Hsiao-Sheng
- Venue
- Experimental Cell Research
- Publisher
- Elsevier BV
- Published
- 1999-05
- Volume
- 248
- Issue
- 2
- Pages
- 589-598
- ISSN
- 0014-4827 (ISSN-L)
- OpenAlex
- W1971946381
- Cited by
- 20
- References
- 48
- FWCI
- 0.80
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Experimental Cell Research (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Keywords
- TNF-α, TNF-alpha cytotoxicity, Raf-1, MAPK signaling, apoptosis, NIH3T3 cells, Rac1, RhoA, Bcl-2, PI3-kinase, cell cycle arrest, AP-1, c-fos, c-Myc
- MeSH terms
- Dose-Response Relationship, Drug, Gene Expression Regulation, Humans, Genes, ras, Transformation, Genetic, Tumor Cells, Cultured, Tumor Necrosis Factor-alpha, Signal Transduction, Proto-Oncogene Proteins c-myc, Proto-Oncogene Proteins c-fos, Apoptosis, Calcium-Calmodulin-Dependent Protein Kinases, ras Proteins, Transcription Factor AP-1, Transgenes, Proto-Oncogene Proteins c-raf, Inhibitory Concentration 50
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