Jian-Ching Wu1, Chiang-Ting Wang2,3, Han-Chun Hung1, Wen-Jeng Wu4,5,6,7,8, Deng-Chyang Wu8,9,10, Min-Chi Chang11, Ping-Jyun Sung12,13, Yu-Wei Chou14, Zhi-Hong Wen15,16, Ming-Hong Tai17,18,19,20. 1. Doctoral Degree Program in Marine Biotechnology, Academia Sinica, National Sun Yat-sen University, Kaohsiung, Taiwan. 2. Department of Urological Surgery, Kaohsiung Armed Force General Hospital, Kaohsiung, Taiwan. 3. Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan. 4. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 5. Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan. 6. Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 7. Department of Urology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan. 8. Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung, Taiwan. 9. Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. 10. Faculty of Medicine, Department of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 11. Division of Colorectal Surgery, Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan. 12. National Museum of Marine Biology and Aquarium, Pingtung, Taiwan. 13. Institute of Marine Biotechnology, National Dong Hwa University, Pingtung, Taiwan. 14. Tissue Bank and BioBank, Kaohsiung Chang Gung Memorial Hospital, Niao-Song District, Kaohsiung, Taiwan. 15. Doctoral Degree Program in Marine Biotechnology, Academia Sinica, National Sun Yat-sen University, Kaohsiung, Taiwan. wzh@mail.nsysu.edu.tw. 16. Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan. wzh@mail.nsysu.edu.tw. 17. Doctoral Degree Program in Marine Biotechnology, Academia Sinica, National Sun Yat-sen University, Kaohsiung, Taiwan. minghongtai@gmail.com. 18. Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung, Taiwan. minghongtai@gmail.com. 19. Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan. minghongtai@gmail.com. 20. Center for Neuroscience, National Sun Yat-sen University, Kaohsiung, Taiwan. minghongtai@gmail.com.
Abstract
BACKGROUND: Prostate cancer is one of the most prevalent cancers in men worldwide. Aberrant activation of c-Met/signal transducer and activator of transcription-3 (STAT3) signaling is involved in prostate carcinogenesis, underscoring the demand for developing c-Met/STAT3-targeting drugs. Thus, we first utilized virtual screening strategy to identify STAT3-inhibiting marine compound, heteronemin, and then validated the STAT3-inhibiting function of heteronemin in prostate cancer cells. METHODS: Human prostate cancer LNCaP, DU145, and PC-3 cell lines were treated with heteronemin for 24 hr, then the cell viability was evaluated by MTT assay. Flow cytometry was performed to analyze the apoptosis in heteronemin-treated cells. Western blot and quantitative real-time PCR were executed to further confirm the c-Met/STAT3 signaling inhibition by heteronemin in DU145 and PC-3 cells. RESULTS: In this study, we employed the virtual screening strategy to identify heteronemin, a spongean sesterterpene, as a potential STAT3 inhibitor from Taiwan marine drugs library. Application of heteronemin potently suppressed the viability and anchorage-independent growth of human prostate cancer cells. Besides, heteronemin induced apoptosis in prostate cancer cells by activation of both intrinsic (caspase-9) and extrinsic (caspase-8) apoptotic pathways. By luciferase assay and expression analysis, it was confirmed that heteronemin inhibited the phosphorylation of c-Met/src/STAT3 signaling axis, STAT3-driven luciferase activities and expression of STAT3-regulated genes including Bcl-xL, Bcl-2, and Cyclin D1. Finally, heteronemin effectively antagonized the hepatocyte growth factor (HGF)-stimulated c-Met/STAT3 activation as well as the proliferation and colonies formation in refractory prostate cancer cells. CONCLUSIONS: These findings suggest that heteronemin may constitute a novel c-Met/STAT3-targeting agent for prostate cancer. Prostate 76:1469-1483, 2016.
BACKGROUND:Prostate cancer is one of the most prevalent cancers in men worldwide. Aberrant activation of c-Met/signal transducer and activator of transcription-3 (STAT3) signaling is involved in prostate carcinogenesis, underscoring the demand for developing c-Met/STAT3-targeting drugs. Thus, we first utilized virtual screening strategy to identify STAT3-inhibiting marine compound, heteronemin, and then validated the STAT3-inhibiting function of heteronemin in prostate cancer cells. METHODS:Humanprostate cancer LNCaP, DU145, and PC-3 cell lines were treated with heteronemin for 24 hr, then the cell viability was evaluated by MTT assay. Flow cytometry was performed to analyze the apoptosis in heteronemin-treated cells. Western blot and quantitative real-time PCR were executed to further confirm the c-Met/STAT3 signaling inhibition by heteronemin in DU145 and PC-3 cells. RESULTS: In this study, we employed the virtual screening strategy to identify heteronemin, a spongean sesterterpene, as a potential STAT3 inhibitor from Taiwan marine drugs library. Application of heteronemin potently suppressed the viability and anchorage-independent growth of humanprostate cancer cells. Besides, heteronemin induced apoptosis in prostate cancer cells by activation of both intrinsic (caspase-9) and extrinsic (caspase-8) apoptotic pathways. By luciferase assay and expression analysis, it was confirmed that heteronemin inhibited the phosphorylation of c-Met/src/STAT3 signaling axis, STAT3-driven luciferase activities and expression of STAT3-regulated genes including Bcl-xL, Bcl-2, and Cyclin D1. Finally, heteronemin effectively antagonized the hepatocyte growth factor (HGF)-stimulated c-Met/STAT3 activation as well as the proliferation and colonies formation in refractory prostate cancer cells. CONCLUSIONS: These findings suggest that heteronemin may constitute a novel c-Met/STAT3-targeting agent for prostate cancer. Prostate 76:1469-1483, 2016.