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Transfection of K-rasAsp12 cDNA markedly elevates IL-1beta- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells.
Oncogene 2003; 22(48):7667-76O

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-rasAsp12 cDNA, the iNOS expression linked to interleukin-1beta (IL-1beta) 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-rasWT. Electrophoretic mobility-shift assays demonstrated that NF-kappaB binding activity induced by IL-1beta or LPS was also increased in K-rasAsp12-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/EBPbeta to a C/EBP-binding consensus site. Furthermore, the anchorage-independent growth of K-rasAsp12-transfected cells was markedly increased by IL-1beta or LPS treatment, and decreased by ONO-1714, an iNOS inhibitor. In addition, tumor growth in nude mice injected with K-rasAsp12-transfected cells was significantly suppressed by NOS inhibition with 50 p.p.m. ONO-1714 or 100 p.p.m. L-NG-nitroarginine methyl ester. These results suggest that an activating mutation of K-ras can markedly enhance the iNOS expression mediated by IL-1beta or LPS, through the activation of promoters on NF-kappaB, C/EBP, and CRE-like sites, and that nitric oxide contributes to the colony formation and tumor growth of K-ras-transformed cells.

Authors+Show Affiliations

Cancer Prevention Basic Research Project, National Cancer Center Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan. mtakahas@gan2.ncc.go.jpNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14576830

Citation

Takahashi, Mami, et al. "Transfection of K-rasAsp12 cDNA Markedly Elevates IL-1beta- and Lipopolysaccharide-mediated Inducible Nitric Oxide Synthase Expression in Rat Intestinal Epithelial Cells." Oncogene, vol. 22, no. 48, 2003, pp. 7667-76.
Takahashi M, Mutoh M, Shoji Y, et al. Transfection of K-rasAsp12 cDNA markedly elevates IL-1beta- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells. Oncogene. 2003;22(48):7667-76.
Takahashi, M., Mutoh, M., Shoji, Y., Kamanaka, Y., Naka, M., Maruyama, T., ... Wakabayashi, K. (2003). Transfection of K-rasAsp12 cDNA markedly elevates IL-1beta- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells. Oncogene, 22(48), pp. 7667-76.
Takahashi M, et al. Transfection of K-rasAsp12 cDNA Markedly Elevates IL-1beta- and Lipopolysaccharide-mediated Inducible Nitric Oxide Synthase Expression in Rat Intestinal Epithelial Cells. Oncogene. 2003 Oct 23;22(48):7667-76. PubMed PMID: 14576830.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transfection of K-rasAsp12 cDNA markedly elevates IL-1beta- and lipopolysaccharide-mediated inducible nitric oxide synthase expression in rat intestinal epithelial cells. AU - Takahashi,Mami, AU - Mutoh,Michihiro, AU - Shoji,Yutaka, AU - Kamanaka,Yoshihisa, AU - Naka,Masao, AU - Maruyama,Takayuki, AU - Sugimura,Takashi, AU - Wakabayashi,Keiji, PY - 2003/10/25/pubmed PY - 2003/12/6/medline PY - 2003/10/25/entrez SP - 7667 EP - 76 JF - Oncogene JO - Oncogene VL - 22 IS - 48 N2 - 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-rasAsp12 cDNA, the iNOS expression linked to interleukin-1beta (IL-1beta) 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-rasWT. Electrophoretic mobility-shift assays demonstrated that NF-kappaB binding activity induced by IL-1beta or LPS was also increased in K-rasAsp12-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/EBPbeta to a C/EBP-binding consensus site. Furthermore, the anchorage-independent growth of K-rasAsp12-transfected cells was markedly increased by IL-1beta or LPS treatment, and decreased by ONO-1714, an iNOS inhibitor. In addition, tumor growth in nude mice injected with K-rasAsp12-transfected cells was significantly suppressed by NOS inhibition with 50 p.p.m. ONO-1714 or 100 p.p.m. L-NG-nitroarginine methyl ester. These results suggest that an activating mutation of K-ras can markedly enhance the iNOS expression mediated by IL-1beta or LPS, through the activation of promoters on NF-kappaB, C/EBP, and CRE-like sites, and that nitric oxide contributes to the colony formation and tumor growth of K-ras-transformed cells. SN - 0950-9232 UR - https://www.unboundmedicine.com/medline/citation/14576830/Transfection_of_K_rasAsp12_cDNA_markedly_elevates_IL_1beta__and_lipopolysaccharide_mediated_inducible_nitric_oxide_synthase_expression_in_rat_intestinal_epithelial_cells_ L2 - http://dx.doi.org/10.1038/sj.onc.1207051 DB - PRIME DP - Unbound Medicine ER -