{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T07:56:21Z","timestamp":1775894181515,"version":"3.50.1"},"reference-count":38,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,3,5]],"date-time":"2025-03-05T00:00:00Z","timestamp":1741132800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Grant-in-Aid for Scientific Research","award":["JP22K09506"],"award-info":[{"award-number":["JP22K09506"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["JP23K08737"],"award-info":[{"award-number":["JP23K08737"]}]},{"name":"Ministry of Education, Science, Sports and Culture and Japan Society for Promotion of Science, Japan","award":["JP22K09506"],"award-info":[{"award-number":["JP22K09506"]}]},{"name":"Ministry of Education, Science, Sports and Culture and Japan Society for Promotion of Science, Japan","award":["JP23K08737"],"award-info":[{"award-number":["JP23K08737"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The liver is the most lethal metastatic site in castration-resistant prostate cancer (CRPC). Overexpression of MET protein has been reported in CRPC, and MET is an important driver gene in androgen-independent CRPC cells. Mouse CRPC cell line CRTC2 was established by subcutaneous injection of hormone-sensitive PC cells (TRAMP-C2) in castrated nude mice. CRCT2\/luc2 cells were injected into the spleen of castrated nude mice, and liver metastasis was confirmed at 2 weeks post-injection. We administered MET inhibitor (MET-I) and HGF activator inhibitor (HGFA-I) to this liver metastasis model and assessed the therapeutic effect. After intrasplenic injection, CRTC2 showed a higher incidence of liver metastasis whereas no metastasis was observed in TRAMP-C2. Microarray analysis revealed increased expression of HGF, MET, and HPN, HGFAC (encoding HGF activating proteases) in liver metastasis. Proliferation of CRCT2 was significantly inhibited by co-administration of MET-I and HGFA-I by in vitro analysis with HGF-enriched condition. In an analysis of the mouse model, the combination-therapy group showed the strongest reduction for liver metastasis. Immunohistochemical staining also revealed the strongest decrease in phosphorylation of MET in the combination-therapy group. Co-culture with HGF-expressed mouse fibroblasts showed attenuation of the inhibitory effect of MET-I; however, additional HGFA-I overcame the resistance. We established an androgen-independent CRPC cell line, CRTC2, and liver metastasis model in mice. Significant effect was confirmed by combined treatment of MET-I and HGFA-I by in vitro and in vivo analysis. The results suggested the importance of combined treatment with both MET- and HGF-targeting agents in the treatment of HGF-enriched conditions including liver metastasis.<\/jats:p>","DOI":"10.3390\/ijms26052308","type":"journal-article","created":{"date-parts":[[2025,3,5]],"date-time":"2025-03-05T05:54:38Z","timestamp":1741154078000},"page":"2308","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Combined Therapy Targeting MET and Pro-HGF Activation Shows Significant Therapeutic Effect Against Liver Metastasis of CRPC"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1123-6815","authenticated-orcid":false,"given":"Shoichi","family":"Kimura","sequence":"first","affiliation":[{"name":"Department of Urology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3757-1504","authenticated-orcid":false,"given":"Satoshi","family":"Iwano","sequence":"additional","affiliation":[{"name":"Institute for Tenure Track Promotion, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Akioka","sequence":"additional","affiliation":[{"name":"Department of Urology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Kuchimaru","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, Jichi Medical University, Tochigi 329-0498, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8210-5460","authenticated-orcid":false,"given":"Makiko","family":"Kawaguchi","sequence":"additional","affiliation":[{"name":"Section of Oncopathology and Morphological Pathology, Department of Pathology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9350-777X","authenticated-orcid":false,"given":"Tsuyoshi","family":"Fukushima","sequence":"additional","affiliation":[{"name":"Section of Oncopathology and Morphological Pathology, Department of Pathology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuichiro","family":"Sato","sequence":"additional","affiliation":[{"name":"Section of Oncopathology and Morphological Pathology, Department of Pathology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9948-0451","authenticated-orcid":false,"given":"Hiroaki","family":"Kataoka","sequence":"additional","affiliation":[{"name":"Organization for Promotion of Research and Industry Academic Regional Collaboration, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshiyuki","family":"Kamoto","sequence":"additional","affiliation":[{"name":"Department of Urology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shoichiro","family":"Mukai","sequence":"additional","affiliation":[{"name":"Department of Urology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsuro","family":"Sawada","sequence":"additional","affiliation":[{"name":"Department of Urology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1683","DOI":"10.1016\/S0140-6736(24)00651-2","article-title":"The Lancet Commission on prostate cancer: Planning for the surge in cases","volume":"403","author":"James","year":"2024","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1002\/pros.24082","article-title":"Androgen receptor signaling-targeted therapy and taxane chemotherapy induce visceral metastasis in castration-resistant prostate cancer","volume":"81","author":"Iwamoto","year":"2021","journal-title":"Prostate"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1016\/j.urology.2009.07.1360","article-title":"Recent time trends in the epidemiology of stage IV prostate cancer in the United States: Analysis of data from the Surveillance, Epidemiology, and End Results Program","volume":"75","author":"Cetin","year":"2010","journal-title":"Urology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1111\/j.1442-2042.2011.02895.x","article-title":"Oncological outcomes of the prostate cancer patients registered in 2004: Report from the Cancer Registration Committee of the JUA","volume":"18","author":"Fujimoto","year":"2011","journal-title":"Int. J. Urol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1652","DOI":"10.1200\/JCO.2015.65.7270","article-title":"Meta-Analysis Evaluating the Impact of Site of Metastasis on Overall Survival in Men With Castration-Resistant Prostate Cancer","volume":"34","author":"Halabi","year":"2016","journal-title":"J. Clin. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"819","DOI":"10.21037\/tau.2020.01.20","article-title":"A case of castration-resistant prostate cancer with liver metastases achieved a complete response by docetaxel chemotherapy","volume":"9","author":"Shirotake","year":"2020","journal-title":"Transl. Androl. Urol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1002\/pros.24400","article-title":"Elevated serum CEA is associated with liver metastasis and distinctive circulating tumor DNA alterations in patients with castration-resistant prostate cancer","volume":"82","author":"Bray","year":"2022","journal-title":"Prostate"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Mukai, S., Yamasaki, K., Fujii, M., Nagai, T., Terada, N., Kataoka, H., and Kamoto, T. (2020). Dysregulation of Type II Transmembrane Serine Proteases and Ligand-Dependent Activation of MET in Urological Cancers. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21082663"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.3390\/cancers6041890","article-title":"Mechanisms of hepatocyte growth factor activation in cancer tissues","volume":"6","author":"Kawaguchi","year":"2014","journal-title":"Cancers"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1111\/pin.12647","article-title":"Hepatocyte growth factor activator inhibitors (HAI-1 and HAI-2): Emerging key players in epithelial integrity and cancer","volume":"68","author":"Kataoka","year":"2018","journal-title":"Pathol. Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"63014","DOI":"10.18632\/oncotarget.18260","article-title":"Targeting the tumor-promoting microenvironment in MET-amplified NSCLC cells with a novel inhibitor of pro-HGF activation","volume":"8","author":"Owusu","year":"2017","journal-title":"Oncotarget"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103234","DOI":"10.1016\/j.critrevonc.2021.103234","article-title":"Combination of HGF\/MET-targeting agents and other therapeutic strategies in cancer","volume":"160","author":"Moosavi","year":"2021","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1007\/s10555-012-9391-7","article-title":"Acquired resistance mechanisms to tyrosine kinase inhibitors in lung cancer with activating epidermal growth factor receptor mutation\u2014Diversity, ductility, and destiny","volume":"31","author":"Suda","year":"2012","journal-title":"Cancer Metastasis Rev."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Owusu, B.Y., Galemmo, R., Janetka, J., and Klampfer, L. (2017). Hepatocyte Growth Factor, a Key Tumor-Promoting Factor in the Tumor Microenvironment. Cancers, 9.","DOI":"10.3390\/cancers9040035"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"6598","DOI":"10.1158\/0008-5472.CAN-14-0761","article-title":"Microenvironment-derived HGF overcomes genetically determined sensitivity to anti-MET drugs","volume":"74","author":"Pennacchietti","year":"2014","journal-title":"Cancer Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1146\/annurev-pathol-052016-100322","article-title":"The Role of Cancer-Associated Fibroblasts and Fibrosis in Liver Cancer","volume":"12","author":"Affo","year":"2017","journal-title":"Annu. Rev. Pathol. Mech. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1038\/s41571-023-00770-1","article-title":"Cholangiocarcinoma\u2014Novel biological insights and therapeutic strategies","volume":"20","author":"Ilyas","year":"2023","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/S2468-1253(23)00111-5","article-title":"Fibroblasts in liver cancer: Functions and therapeutic translation","volume":"8","author":"Affo","year":"2023","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"866","DOI":"10.1016\/j.ccell.2021.03.012","article-title":"Promotion of cholangiocarcinoma growth by diverse cancer-associated fibroblast subpopulations","volume":"39","author":"Affo","year":"2021","journal-title":"Cancer Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1002\/pros.20714","article-title":"Broadening of transgenic adenocarcinoma of the mouse prostate (TRAMP) model to represent late stage androgen depletion independent cancer","volume":"68","author":"Jeet","year":"2008","journal-title":"Prostate"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1002\/pros.24055","article-title":"A hemi-spleen injection model of liver metastasis for prostate cancer","volume":"80","author":"Simons","year":"2020","journal-title":"Prostate"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1152\/physrev.00048.2019","article-title":"Diversity and biology of cancerassociated fibroblasts","volume":"101","author":"Biffi","year":"2021","journal-title":"Physiol. Rev."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Janetka, W.J., and Benson, M.R. (2018). Extracellular Targeting of Cell Signaling in Cancer, Wiley. Strategies Directed at MET and RON Receptor Tyrosine Kinase Pathways.","DOI":"10.1002\/9781119300229"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1016\/S0090-4295(02)01954-4","article-title":"High expression of the Met receptor in prostate cancer metastasis to bone","volume":"60","author":"Knudsen","year":"2002","journal-title":"Urology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"31029","DOI":"10.18632\/oncotarget.8829","article-title":"MET expression during prostate cancer progression","volume":"7","author":"Verhoef","year":"2016","journal-title":"Oncotarget"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/S0002-9440(10)63318-4","article-title":"Phenotypic switch from paracrine to autocrine role of hepatocyte growth factor in an androgen-independent human prostatic carcinoma cell line, CWR22R","volume":"165","author":"Nakashiro","year":"2004","journal-title":"Am. J. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.2217\/fon-2021-1096","article-title":"A phase III, randomized, open-label study (CONTACT-02) of cabozantinib plus atezolizumab versus second novel hormone therapy in patients with metastatic castration-resistant prostate cancer","volume":"18","author":"Agarwal","year":"2022","journal-title":"Future Oncol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1007\/s13577-015-0123-5","article-title":"Plasma macrophage-stimulating protein and hepatocyte growth factor levels are associated with prostate cancer progression","volume":"29","author":"Sugie","year":"2016","journal-title":"Hum. Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"686","DOI":"10.1016\/j.eururo.2005.05.020","article-title":"Serum hepatocyte growth factor activator (HGFA) in benign prostatic hyperplasia and prostate cancer","volume":"48","author":"Nagakawa","year":"2005","journal-title":"Eur. Urol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Yao, J.F., Li, X.J., Yan, L.K., He, S., Zheng, J.B., Wang, X.R., Zhou, P.H., Zhang, L., Wei, G.B., and Sun, X.J. (2019). Role of HGF\/c-Met in the treatment of colorectal cancer with liver metastasis. J. Biochem. Mol. Toxicol., 33.","DOI":"10.1002\/jbt.22316"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1159\/000065477","article-title":"c-Met expression in gastric cancer with liver metastasis","volume":"63","author":"Amemiya","year":"2002","journal-title":"Oncology"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Elliott, V.A., Rychahou, P., Zaytseva, Y.Y., and Evers, B.M. (2014). Activation of c-Met and upregulation of CD44 expression are associated with the metastatic phenotype in the colorectal cancer liver metastasis model. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0097432"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"15016","DOI":"10.1038\/ncomms15016","article-title":"Exosome-delivered EGFR regulates liver microenvironment to promote gastric cancer liver metastasis","volume":"8","author":"Zhang","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1038\/sj.cgt.7700705","article-title":"Hepatic gene expression of NK4, an HGF-antagonist\/angiogenesis inhibitor, suppresses liver metastasis and invasive growth of colon cancer in mice","volume":"11","author":"Wen","year":"2004","journal-title":"Cancer Gene Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"29492","DOI":"10.18632\/oncotarget.8785","article-title":"Inhibition of pro-HGF activation by SRI31215, a novel approach to block oncogenic HGF\/MET signaling","volume":"7","author":"Owusu","year":"2016","journal-title":"Oncotarget"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1002\/ijc.31550","article-title":"Targeting the MET oncogene by concomitant inhibition of receptor and ligand via an antibody-\u201cdecoy\u201d strategy","volume":"143","author":"Basilico","year":"2018","journal-title":"Int. J. Cancer"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"So, S., Park, Y., Kang, S.S., Han, J., Sunwoo, J.H., Lee, W., Kim, J., Ye, E.A., Kim, J.Y., and Tchah, H. (2022). Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24010701"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1021\/acsmedchemlett.5b00357","article-title":"Design and Synthesis of Nonpeptide Inhibitors of Hepatocyte Growth Factor Activation","volume":"7","author":"Venukadasula","year":"2016","journal-title":"ACS Med. Chem. Lett."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/5\/2308\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:47:25Z","timestamp":1760028445000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/5\/2308"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,5]]},"references-count":38,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["ijms26052308"],"URL":"https:\/\/doi.org\/10.3390\/ijms26052308","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,5]]}}}