Transfection of k-rasasp12 cdna markedly elevates il-1β- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells

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,

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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