Ras up-regulation of cyclooxygenase-2.
Oncogenic mutations in Ras (H-Ras, N-Ras, and K-Ras) are found in a wide variety of human malignancies, including adenocarcinomas of the colon, where K-Ras mutations often occur early in tumor development and strongly correlate with the transition to invasive adenocarcinoma. Our laboratory is interested in examining the interaction between Ras signaling and up-regulation of cyclooxygenase-2 (COX-2), a key regulator of prostaglandin biosynthesis. Our studies demonstrate that the Ras oncoprotein can regulate transcriptional activation and stabilization of COX-2 expression by several mechanisms. In this chapter we have outlined protocols and experimental approaches used in our laboratory to measure H-Ras up-regulation of COX-2 expression and to elaborate on more recent techniques that illustrate the importance of activation of Ras by prostaglandin E2 (PGE(2)). These methods have facilitated our understanding of the mechanisms by which the COX-2-derived PGE(2) and Ras activation of the mitogen-activated protein kinase (MAPK) signaling promotes oncogenic transformation. In light of the critical roles of both COX-2 and Ras signaling in carcinogenesis, our understanding of the complete signaling nuances between different isoforms of Ras on activation of COX-2, as well as understanding the novel mechanism whereby COX-2-derived PGE(2) constitutively activates Ras, will potentially aid in the identification of new targets for cancer therapy.
Backlund
Michael G
MG
Department of Medicine, Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Mann
Jason R
JR
Wang
Dingzhi
D
Dubois
Raymond N
RN
D000818:Animals; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D015232:Dinoprostone / Q000502:physiology; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D020935:MAP Kinase Signaling System; D051381:Rats; D015854:Up-Regulation; D018631:ras Proteins / Q000502:physiology*
D051546:Cyclooxygenase 2; D018631:ras Proteins; D015232:Dinoprostone