Department of Precision Medicine, School of Medicine, University of Campania 'L. Vanvitelli', Via L. De Crecchio 7, 80138, Naples, Italy.
Department of Pharmacy, University Federico II of Naples, Via D. Montesano 49, 80131, Naples, Italy.
Sci Rep. 2021 Dec 1;11(1):23232. doi: 10.1038/s41598-021-02675-4.
Transient receptor potential melastatin-8 (TRPM8) represents an emerging target in prostate cancer, although its mechanism of action remains unclear. Here, we have characterized and investigated the effects of TRPM8 modulators in prostate cancer aggressiveness disclosing the molecular mechanism underlying their biological activity. Patch-clamp and calcium fluorometric assays were used to characterize the synthesized compounds. Androgen-stimulated prostate cancer-derived cells were challenged with the compounds and the DNA synthesis was investigated in a preliminary screening. The most effective compounds were then employed to inhibit the pro-metastatic behavior of in various PC-derived cells, at different degree of malignancy. The effect of the compounds was then assayed in prostate cancer cell-derived 3D model and the molecular targets of selected compounds were lastly identified using transcriptional and non-transcriptional reporter assays. TRPM8 antagonists inhibit the androgen-dependent prostate cancer cell proliferation, migration and invasiveness. They are highly effective in reverting the androgen-induced increase in prostate cancer cell spheroid size. The compounds also revert the proliferation of castrate-resistant prostate cancer cells, provided they express the androgen receptor. In contrast, no effects were recorded in prostate cancer cells devoid of the receptor. Selected antagonists interfere in non-genomic androgen action and abolish the androgen-induced androgen receptor/TRPM8 complex assembly as well as the increase in intracellular calcium levels in prostate cancer cells. Our results shed light in the processes controlling prostate cancer progression and make the transient receptor potential melastatin-8 as a 'druggable' target in the androgen receptor-expressing prostate cancers.
瞬时受体电位 melastatin-8（TRPM8）是前列腺癌的一个新兴靶点，尽管其作用机制尚不清楚。在这里，我们对 TRPM8 调节剂在前列腺癌侵袭性中的作用进行了表征和研究，揭示了其生物学活性的潜在分子机制。我们使用膜片钳和钙荧光检测法来表征合成的化合物。用这些化合物处理雄激素刺激的前列腺癌细胞，并在初步筛选中研究 DNA 合成。然后，我们使用最有效的化合物来抑制各种前列腺癌细胞来源的细胞的促转移行为，其恶性程度不同。然后在前列腺癌细胞衍生的 3D 模型中检测化合物的作用，最后使用转录和非转录报告基因检测来鉴定选定化合物的分子靶标。TRPM8 拮抗剂抑制雄激素依赖性前列腺癌细胞的增殖、迁移和侵袭。它们在逆转雄激素诱导的前列腺癌细胞球体大小增加方面非常有效。这些化合物还可以逆转去势抵抗性前列腺癌细胞的增殖，只要它们表达雄激素受体。相比之下，在没有受体的前列腺癌细胞中没有记录到这些作用。选定的拮抗剂干扰非基因组雄激素作用，并消除雄激素诱导的雄激素受体/TRPM8 复合物组装以及前列腺癌细胞内钙水平的增加。我们的结果阐明了控制前列腺癌进展的过程，并使瞬时受体电位 melastatin-8 成为表达雄激素受体的前列腺癌的“可用药”靶点。
