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Dual roles of AMAP1 in the transcriptional regulation and intracellular trafficking of carbonic anhydrase IX
Title: | Dual roles of AMAP1 in the transcriptional regulation and intracellular trafficking of carbonic anhydrase IX |
Authors: | Horikawa, Mei Browse this author | Sabe, Hisataka Browse this author | Onodera, Yasuhito Browse this author →KAKEN DB |
Keywords: | CA9 | ASAP1 | Protein kinase D2 | PIAS3 | HIF1A |
Issue Date: | Jan-2022 |
Publisher: | Elsevier |
Journal Title: | Translational Oncology |
Volume: | 15 |
Issue: | 1 |
Start Page: | 101258 |
Publisher DOI: | 10.1016/j.tranon.2021.101258 |
Abstract: | Background: The cell-surface enzyme carbonic anhydrase IX (CAIX/CA9) promotes tumor growth, survival, invasion, and metastasis, mainly via its pH-regulating functions. Owing to its tumor-specific expression, CAIXtargeting antibodies/chemicals are utilized for therapeutic and diagnostic purposes. However, mechanisms of CAIX trafficking, which affects such CAIX-targeting modalities remain unclear. In this study, roles of the AMAP1PRKD2 pathway, which mediates integrin recycling of invasive cancer cells, in CAIX trafficking were investigated.Methods: Using highly invasive MDA-MB-231 breast cancer cells, the physical association and colocalization of endogenous proteins were analyzed by immunoprecipitation and immunofluorescence, protein/mRNA levels were quantified by western blotting/qPCR, and cell-surface transport and intracellular/extracellular pH regulation were measured by biotin-labeling and fluorescent dye-based assays, respectively. The correlation between mRNA levels and patients' prognoses was analyzed using a TCGA breast cancer dataset.Results: AMAP1 associated with the CAIX protein complex, and they colocalized at the plasma membrane and tubulovesicular structures. AMAP1 knockdown reduced total/surface CAIX, induced its lysosomal accumulation and degradation, and affected intracellular/extracellular pH. PRKD2 knockdown excluded AMAP1 from the CAIX complex and reduced total CAIX in a lysosome-dependent manner. Unexpectedly, AMAP1 knockdown also reduced CAIX mRNA. AMAP1 interacted with PIAS3, which stabilizes HIF-1 alpha, a transcriptional regulator of CA9. AMAP1 knockdown inhibited the PIAS3-HIF-1 alpha interaction and destabilized the HIF-1 alpha protein. High-ASAP1 (AMAP1-encoding gene) together with high-PIAS3 correlated with high-CA9 and an unfavorable prognosis in breast cancer. Conclusion: The AMAP1-PRKD2 pathway regulates CAIX trafficking, and modulates its total/surface expression. The AMAP1-PIAS3 interaction augments CA9 transcription by stabilizing HIF-1 alpha, presumably contributing to an unfavorable prognosis. |
Type: | article |
URI: | http://hdl.handle.net/2115/85688 |
Appears in Collections: | 医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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