背景：瞬时受体电位香草受体亚家族成员6（TRPV6）是一种高度特异性的钙离子通道，已经证实在钙离子平衡方面发挥着重要作用，并且在前列腺癌的生长、细胞存活和药物抵抗等方面在体外和体内中参与其中。其作用及相应的钙离子依赖通路主要在激素依赖型人类前列腺癌细胞系中进行研究，通常用作早期前列腺癌模型。本研究的目标是通过使用抗去势型前列腺癌细胞系，描述TRPV6特异表型及其参与的信号通路。方法：使用RNA测序（RNA-seq）研究由TRPV6影响的基因表达，使用PC3Mtrpv6-/-与PC3Mtrpv6+/+及其衍生物PC3M-luc-C6trpv6+/+细胞株在其本身和TRPV6过量表达形式中进行研究。除了全细胞RNA测序外，还使用免疫印迹、定量PCR和钙成像等方法验证了trpv6基因状态和功能后果，包括trpv6-/-和TRPV6过表达细胞系。结果：使用免疫印迹和定量PCR验证了trpv6-/-状态，并使用钙成像显示了trpv6基因缺失或TRPV6过表达的功能后果。RNA测序分析表明，作为钙信号的关键调节者，钙通道TRPV6显著影响与癌症进展相关的基因表达，例如细胞周期调节、趋化性、迁移、侵袭、凋亡、铁死亡以及药物抵抗和细胞外基质重组。结论：我们的数据表明，trpv6基因参与并调节了与抗去势型前列腺癌细胞的肿瘤进展和药物抵抗相关的多个通路。版权所有© 2023 Cordier, Haustrate, Prevarskaya and Lehen’kyi.

Background: 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 in vitro and in vivo 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. Methods: RNA sequencing (RNA-seq) was used to study the gene expression impacted by TRPV6 using PC3Mtrpv6-/- versus PC3Mtrpv6+/+ and its derivative PC3M-luc-C6trpv6+/+ 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 trpv6 gene status and functional consequences, in both trpv6 -/- and TRPV6 overexpression cell lines. Results: trpv6 -/- status was validated using both immunoblotting and quantitative PCR, and the functional consequences of either trpv6 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. Conclusion: Our data suggest that the trpv6 gene is involved in and regulates multiple pathways related to tumor progression and drug resistance in castration-resistant prostate cancer cells.Copyright © 2023 Cordier, Haustrate, Prevarskaya and Lehen’kyi.