AUTHOR=Cordier Clément , Haustrate Aurélien , Prevarskaya Natalia , Lehen’kyi V’yacheslav 

TITLE=Characterization of the TRPV6 calcium channel-specific phenotype by RNA-seq in castration-resistant human prostate cancer cells

JOURNAL=Frontiers in Genetics

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2023.1215645

DOI=10.3389/fgene.2023.1215645

ISSN=1664-8021

ABSTRACT=<p><bold>Background:</bold> Transient receptor potential vanilloid subfamily member 6 (TRPV6), a highly calcium-selective channel, has been shown to play a significant role in calcium homeostasis and to participate both <italic>in vitro</italic> and <italic>in vivo</italic> in growth, cell survival, and drug resistance of prostate cancer. Its role and the corresponding calcium-dependent pathways were mainly studied in hormone-dependent human prostate cancer cell lines, often used as a model of early-stage prostate cancers. The goal of the present study was to describe the TRPV6-specific phenotype and signaling pathways it is involved in, using castration-resistant prostate cancer cell lines.</p><p><bold>Methods:</bold> RNA sequencing (RNA-seq) was used to study the gene expression impacted by TRPV6 using PC3M<sup><italic>trpv6−/−</italic></sup><italic>versus</italic> PC3M<sup><italic>trpv6+/+</italic></sup> and its derivative PC3M-luc-C6<sup><italic>trpv6+/+</italic></sup> cell line in its native and TRPV6 overexpressed form. In addition to the whole-cell RNA sequencing, immunoblotting, quantitative PCR, and calcium imaging were used to validate <italic>trpv6</italic> gene status and functional consequences, in both <italic>trpv6</italic><sup><italic>-/-</italic></sup> and TRPV6 overexpression cell lines.</p><p><bold>Results:</bold><italic>trpv6</italic><sup>-/-</sup> status was validated using both immunoblotting and quantitative PCR, and the functional consequences of either <italic>trpv6</italic> gene deletion or TRPV6 overexpression were shown using calcium imaging. RNA-seq analysis demonstrated that the calcium channel TRPV6, being a crucial player of calcium signaling, significantly impacts the expression of genes involved in cancer progression, such as cell cycle regulation, chemotaxis, migration, invasion, apoptosis, ferroptosis as well as drug resistance, and extracellular matrix (ECM) re-organization.</p><p><bold>Conclusion:</bold> Our data suggest that the <italic>trpv6</italic> gene is involved in and regulates multiple pathways related to tumor progression and drug resistance in castration-resistant prostate cancer cells.</p>