MCTP1促進攝護腺惡性腫瘤經由ZBTB46/FOXA2/HIF1A 路徑強化SNAI1誘導之神經內分泌分化及上皮細胞間質化表現
何岩1、劉晏年2、陳威宇3,4、葉秀蓮5、陳威豪2、江國慶2、陳子晴6、蕭宏昇7、黄教悌8、溫玉清1,9,10
臺北市立萬芳醫院-委託財團法人臺北醫學大學辦理泌尿科1、臺北醫學大學癌症生物學與藥物研發研究所2、臺北市立萬芳醫院-委託財團法人臺北醫學大學辦理病理科3、臺北醫學大學醫學院醫學系病理學科4、國立清華大學資訊系統與應用研究所5、臺北醫學大學藥學系臨床藥學組6、中央研究院基因體研究中心7、美國杜克大學醫學中心病理科8、臺北醫學大學醫學院醫學系泌尿學科9、臺北醫學大學泌尿腎臟研究中心10
MCTP1 promotes SNAI1-driven neuroendocrine differentiation and epithelial-to-mesenchymal transition of prostate cancer enhancement by ZBTB46/FOXA2/HIF1A
Yen Ho1, Yen-Nien Liu2, Wei-Yu Chen3,4, Hsiu-Lien Yeh5, Wei-Hao Chen2, Kuo-Ching Jiang2, Zi-Qing Chen6, Michael Hsiao7, Jiaoti Huang 8, Yu-Ching Wen 1,9,10
Department of Urology, Wan Fang Hospital, Taipei Medical University1, Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University2, Department of Pathology, Wan Fang Hospital, Taipei Medical University3, Department of Pathology, School of Medicine, College of Medicine, Taipei Medical University4, Institute of Information System and Applications, National Tsing Hua University5, Division of Clinical Pharmacy, School of Pharmacy, Taipei Medical University6, Genomics Research Center, Academia Sinica7, Department of Pathology, Duke University Medical Center, Durham, USA8, Department of Urology, School of Medicine, College of Medicine, Taipei Medical University9, TMU Research Center of Urology and Kidney, Taipei Medical University10
Background: Prostate cancer (PCa) patients with bone metastases frequently exhibit abnormal calcium homeostasis. However, the molecular mechanisms underlying bone metastasis mediated by activated calcium transport signaling remain unclear. Our results showed that androgen deprivation therapy (ADT) of PCa upregulates a calcium sensor transmembrane protein, the multiple C2 domains transmembrane protein 1(MCTP1), which is upregulated by hypoxia-induced ZBTB46/FOXA2/HIF1A signaling and is associated with SNAI1-driven neuroendocrine (NE) differentiation (NED) and epithelial-to-mesenchymal transition (EMT) in PCa.
Methods: Hypoxia induced interactions between ZBTB46, HIF1A, and FOXA2 were confirmed by immunoprecipitation (IP)-Western Blot (WB). Correlation between MCTP1 and ZBTB46/FOXA2/HIF1A complexes was analyzed by immunohistochemical (IHC) staining in consecutive tissue sections of PCa tissue microarray (TMA) comprising primary PCa and small cell PCa (SCPC) samples. The regulatory network between MCTP1 and the hypoxia-induced ZBTB46/FOXA2/HIF1A axis was investigated by chromatin IP (ChIP)-Seq analysis, ChIP assay, and promoter reporter assays. The effect of MCTP1 on SNAI1 expression driving NED and EMT in PCa was identified in PCa cell lines using mimic hypoxia, ADT, or long-term androgen receptor (AR) antagonist culture systems. Activation of the MCTP1-mediated calcium-related pathway induces SNAI1-driven NED and aggressive progression of PCa under hypoxic conditions was examined by Fluo-8 AM staining combined with flow cytometry, migration, invasion, and xenograft model validation.
Results: MCTP1 is highly expressed in advanced PCa tissues and is associated with ZBTB46/FOXA2/HIF1A abundance. Upregulation of MCTP1 enhances SNAI1-mediated cell migration and EMT responses and is associated with hypoxia-driven NED in PCa after ADT. Mechanistically, hypoxia enhances the MCTP1-mediated calcium/AKT pathway to stabilize SNAI1 in PCa to promote EMT and NED by increasing SNAI1-driven expression of EMT and NE markers. MCTP1 knockdown abrogated hypoxia-enhanced EMT and NED by reducing calcium/AKT pathway-driven SNAI1 expression.
Conclusion: Our study shows the molecular mechanism by which hypoxia-associated ZBTB46/FOXA2/HIF1A transcription factor interaction upregulates the calcium sensor transmembrane protein MCTP1 in ADT- resistant PCa, and explores how MCTP1 promotes NED and EMT in PCa by upregulating SNAI1-driven EMT and NE markers.