Mp85-04 de novo gtp synthesis is critical for cell migration through impdh2 in renal cell carcinoma

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology II1 Apr 2016MP85-04 DE NOVO GTP SYNTHESIS IS CRITICAL FOR CELL MIGRATION THROUGH IMPDH2 IN RENAL CELL CARCINOMA Hirofumi Yoshino, Hideki Hideki, Masayuki Nakagawa, and Atsuo Sasaki Hirofumi YoshinoHirofumi Yoshino More articles by this author , Hideki HidekiHideki Hideki More articles by this author , Masayuki NakagawaMasayuki Nakagawa More articles by this author , and Atsuo SasakiAtsuo Sasaki More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2270AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES In the strategy of cancer treatment, it is important to target cancer specific pathway on survival and proliferation of cancer cells. In cells, there are two distinct pathways to generate Guanosine-5'-triphosphate (GTP): salvage and de novo pathway. The salvage pathway is an energy efficient pathway that utilizes assembled parts of nucleotides, which are formed during degradation of DNA and RNA. On the other hand, de novo pathway is an energy consuming, multi-step pathway that uses glucose as the starting material for the formation of GTP. Interestingly, the energy consuming de novo pathway is mainly employed by rapidly proliferating cancer cells, while the energy-efficient salvage pathway is predominantly used by non-cancerous cells. Then we focused on inosine 5'-monophosphate dehydrogenase 2 (IMPDH2) which is a enzyme on GTP de novo pathway. The aim of the present study is to find a novel molecular network involved in renal cell carcinoma (RCC) invasion and metastasis through investigating the function of GTP synthesis. METHODS Expression levels of IMPDH2 were examined by immunohistochemistry (IHC) in clinical RCC tissue array (70 RCCs and 10 normal renal tissues). GTP levels were measured by high performance liquid chromatography (HPLC) in RCC cell lines (786-o and A498) and normal mouse primary cells treated by IMPDH2 inhibitor (mycophenolic acid (MPA)). The functional studies of IMPDH2 were performed to investigate cell migration by RCC cell lines. Fluorescence resonance energy transfer (FRET) analyses were performed to monitor RhoA activity with RhoA biosensor (W47). GTP pull down assay was performed to monitor GTP- bound form of Rho. RESULTS The expression levels of IMPDH2 were significantly up-regulated in RCC tissues in IHC. GTP levels were significantly reduced by MPA in RCC cell lines, on the other hand those in primary cell were not effected. GTP reduction inhibited cell migration in RCC cell lines. FRET analysis showed that Rho activation signals were significantly reduced by MPA. In addition, GTP pull down assay showed that active form of Rho was inhibited by MPA. CONCLUSIONS IMPDH2 was up-regulated in RCCs, and reduction of IMPDH2 inhibited cell migration through GTP down-regulation. FRET and GTP pull down assay indicated that Rho inactivation might be associated with inhibition of cell migration. Our data suggest that IMPDH2 could be a new molecular maker for RCC. In addition, control of GTP could be a new therapeutic target for RCC metastasis and invasion through IMPDH2 regulation. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1099 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Hirofumi Yoshino More articles by this author Hideki Hideki More articles by this author Masayuki Nakagawa More articles by this author Atsuo Sasaki More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...