Determination of Six Transmembrane Protein of Prostate 2 Gene Expression and Intracellular Localization in Prostate Cancer

Abstract

Objective: In this study, we aimed to determine the relationship between the RNA and protein expression profile of six transmembrane protein of prostate 2 (STAMP2) gene and androgen and the intracellular localization of STAMP2. Materials and Methods: RNA and protein were obtained from androgen treated lymph node carcinoma of the prostate (LNCaP) cells, untreated LNCaP cells, DU145 cells with no androgen receptor, and STAMP2 transfected COS-7 cells. The expression profile of STAMP2 gene and the effect of androgenes on the expression was shown in RNA and protein levels by using Northern and Western blotting methods. In addition, intracellular localization of the naturally synthesized STAMP2 protein and the transfected STAMP2 protein in COS-7 cells after androgen administration in both LNCaP cells was determined by immunofluorescence microscopy. Results: We found that the RNA and protein expression of STAMP2 gene in LNCaP cells are regulated by androgenes, the power of expression is increased with the duration of androgen treatment and there is no STAMP2 expression in DU145 cells which has no androgen receptor. As a result of the immunofluorescence microscopy study we observed that STAMP2 protein was localized at golgi complex and cell membrane. Conclusion:In conclusion, we have demonstrated that STAMP2 may play an important role in the pathogenesis of the prostate cancer and in the androgen-dependent androgen-independent staging of prostate cancer. In addition, STAMP2 protein, which is localized in the intracellular golgi complex and cell membrane, may be a new target molecule for prostate cancer diagnosis and treatment. Keywords: Prostate, prostate cancer, six transmembrane protein of prostate 2 Amaç: Çalışmamızda six transmembrane protein of prostate 2 (STAMP2) geninin RNA ve protein ekspresyon profilinin androjenlerle ilişkisinin ve STAMP2'nin hücre içi lokalizasyonunun belirlenmesini amaçladık. Gereç ve Yöntem: RNA ve proteinler androjen uygulanmış ve uygulanmamış prostat kanserinin lenf nodu (LNCaP) hücrelerinden, androjen reseptörü içermeyen DU145 hücrelerinden ve STAMP2 transfekte edilmiş COS-7 hücrelerinden elde edildi. STAMP2 geninin ekspresyon profili ve androjen uygulamasının STAMP2 RNA ve protein düzeylerine etkisi northern blot ve western blot yöntemleri ile gösterildi. Ek olarak hem LNCaP hücrelerinde androjen uygulaması sonrası doğal olarak sentezlenen STAMP2 proteininin hem de COS-7 hücrelerindeki transfekte STAMP2 proteininin hücre içi lokalizasyonu immünfloresan mikroskopi ile belirlendi. Bulgular: LNCaP hücrelerinde STAMP2 geninin RNA ve protein ekspresyonunun androjenlerle düzenlendiği ve androjen uygulama süresi arttıkça ekspresyon düzeyinin arttığı, androjen reseptörü içermeyen DU145 hücrelerinde STAMP2 ekspresyonunun olmadığı saptandı. İmmünfloresan mikroskopi incelemesi sonucunda STAMP2 proteininin hücre içinde golgi cisimciğinde ve hücre membranında lokalize olduğu gözlendi. Sonuç: STAMP2 geni, prostat kanseri patogenezinde ve prostat kanserinin androjen bağımlı evreden androjen bağımsız e...