Transfection of k-rasasp12 cdna markedly elevates il-1β- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells
Transfection of k-rasasp12 cdna markedly elevates il-1β- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells"
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ABSTRACT Activating mutations of K-_ras_ are frequent in colon tumors and aberrant crypt foci, and may play important roles in colon carcinogenesis. Here, we investigated the effects of a
K-_ras_ codon 12 mutation on inducible nitric oxide synthase (iNOS) expression. When rat intestinal epithelial cells (IEC-6) were transfected with K-_ras_Asp12 cDNA, the iNOS expression
linked to interleukin-1_β_ (IL-1_β_) or lipopolysaccharide (LPS) treatment was markedly increased and prolonged. In contrast, it was only very faint and transient in cells transfected with
the control vector or K-_ras_WT. Electrophoretic mobility-shift assays demonstrated that NF-_κ_B binding activity induced by IL-1_β_ or LPS was also increased in K-_ras_Asp12-transfected
cells, along with the binding of CREB-1, CREM-1, ATF-1, ATF-2, and Jun D to a cAMP-responsive element (CRE)-like site and the binding of C/EBP_β_ to a C/EBP-binding consensus site.
Furthermore, the anchorage-independent growth of K-_ras_Asp12-transfected cells was markedly increased by IL-1_β_ or LPS treatment, and decreased by ONO-1714, an iNOS inhibitor. In addition,
tumor growth in nude mice injected with K-_ras_Asp12-transfected cells was significantly suppressed by NOS inhibition with 50 p.p.m. ONO-1714 or 100 p.p.m. L-_N_G-nitroarginine methyl
ester. These results suggest that an activating mutation of K-_ras_ can markedly enhance the iNOS expression mediated by IL-1_β_ or LPS, through the activation of promoters on NF-_κ_B,
C/EBP, and CRE-like sites, and that nitric oxide contributes to the colony formation and tumor growth of K-_ras_-transformed cells. Access through your institution Buy or subscribe This is a
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Download references ACKNOWLEDGEMENTS This work was supported in part by a grant-in-aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS), grants-in-aid for
Cancer Research and for the Second-Term Comprehensive 10-Year Strategy for Cancer Control from the Ministry of Health, Labour and Welfare of Japan, and a grant for the Research on Advanced
Medical Technology from the Ministry of Health, Labour, and Welfare of Japan. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Cancer Prevention Basic Research Project, National Cancer Center
Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo, 104-0045, Japan Mami Takahashi, Michihiro Mutoh, Yutaka Shoji, Takashi Sugimura & Keiji Wakabayashi * Minase Research Institute,
Ono Pharmaceutical Co. Ltd, 1-1, Sakurai 3-chome, Shimamoto-cho, Mishima-gunn, Osaka, 618-8585, Japan Yoshihisa Kamanaka, Masao Naka & Takayuki Maruyama Authors * Mami Takahashi View
author publications You can also search for this author inPubMed Google Scholar * Michihiro Mutoh View author publications You can also search for this author inPubMed Google Scholar *
Yutaka Shoji View author publications You can also search for this author inPubMed Google Scholar * Yoshihisa Kamanaka View author publications You can also search for this author inPubMed
Google Scholar * Masao Naka View author publications You can also search for this author inPubMed Google Scholar * Takayuki Maruyama View author publications You can also search for this
author inPubMed Google Scholar * Takashi Sugimura View author publications You can also search for this author inPubMed Google Scholar * Keiji Wakabayashi View author publications You can
also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Mami Takahashi. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS
ARTICLE Takahashi, M., Mutoh, M., Shoji, Y. _et al._ Transfection of K-_ras_Asp12 cDNA markedly elevates IL-1_β_- and lipopolysaccharide-mediated inducible nitric oxide synthase expression
in rat intestinal epithelial cells. _Oncogene_ 22, 7667–7676 (2003). https://doi.org/10.1038/sj.onc.1207051 Download citation * Received: 31 October 2002 * Revised: 02 June 2003 * Accepted:
31 July 2003 * Published: 23 October 2003 * Issue Date: 23 October 2003 * DOI: https://doi.org/10.1038/sj.onc.1207051 SHARE THIS ARTICLE Anyone you share the following link with will be able
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initiative KEYWORDS * inducible nitric oxide synthase * K-_ras_ * rat * IL-1_β_ * lipopolysaccharide
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