Since its first identification in prostate cancers and prostate tissues, transient receptor potential melastatin-subfamily member 8 (TRPM8) is subsequently found to be overexpressed in a wide range of cancers and is shown to be implicated in tumorigenesis and tumor progression. Here, we used N-(3-aminopropyl)-2-[(3-methylphenyl) methoxy] -N-(2-thienylmethyl) benzamide hydrochloride (AMTB), a specific TRPM8 antagonist, to explore its antitumoral effect on osteosarcoma. We find that AMTB suppresses osteosarcoma cell proliferation, metastasis and induces cellular apoptosis. Xenograft model in nude mice experiments also define that AMTB can increase the sensitivity of tumor cells to cisplatin, the cytotoxic chemotherapeutic regimens in treating osteosarcoma. Molecularly, AMTB specifically antagonizes TRPM8 which is upregulated in osteosarcoma and its expression level in osteosarcoma tissues is negatively related to patients’ prognosis. Finally, RNA sequencing analysis was performed to explore the mechanism underlying the antitumoral effect of AMTB on osteosarcoma cells and the results prove that AMTB suppresses the Transforming Growth Factor β (TGFβ) signaling pathway. Our study provides evidence that TRPM8 could be a potential therapeutic target and AMTB can suppress growth and metastasis of osteosarcoma cells through repressing the TGFβ signaling pathway and increase the sensitivity of tumor cells to cisplatin.

自从它在前列腺癌和前列腺组织中首次被发现后，瞬时受体电位 melastatin 亚家族成员 8 (TRPM8) 随后被发现在多种癌症中过度表达，并显示与肿瘤发生和肿瘤进展有关。在这里，我们使用 N-(3-氨基丙基)-2-[(3-甲基苯基) 甲氧基]-N-(2-噻吩基甲基) 苯甲酰胺盐酸盐 (AMTB)，一种特定的 TRPM8 拮抗剂，来探索其对骨肉瘤的抗肿瘤作用。我们发现 AMTB 抑制骨肉瘤细胞增殖、转移并诱导细胞凋亡。裸鼠异种移植模型实验也确定AMTB可以增加肿瘤细胞对顺铂的敏感性，顺铂是治疗骨肉瘤的细胞毒性化疗方案。分子上，AMTB特异性拮抗在骨肉瘤中上调的TRPM8，其在骨肉瘤组织中的表达水平与患者的预后呈负相关。最后，通过RNA测序分析AMTB对骨肉瘤细胞抗肿瘤作用的机制，结果证明AMTB抑制转化生长因子β（TGFβ）信号通路。我们的研究提供的证据表明 TRPM8 可能是一个潜在的治疗靶点，AMTB 可以通过抑制 TGFβ 信号通路抑制骨肉瘤细胞的生长和转移，并增加肿瘤细胞对顺铂的敏感性。进行RNA测序分析以探索AMTB对骨肉瘤细胞的抗肿瘤作用的机制，结果证明AMTB抑制转化生长因子β（TGFβ）信号通路。我们的研究提供的证据表明 TRPM8 可能是一个潜在的治疗靶点，AMTB 可以通过抑制 TGFβ 信号通路抑制骨肉瘤细胞的生长和转移，并增加肿瘤细胞对顺铂的敏感性。进行RNA测序分析以探索AMTB对骨肉瘤细胞的抗肿瘤作用的机制，结果证明AMTB抑制转化生长因子β（TGFβ）信号通路。我们的研究提供的证据表明 TRPM8 可能是一个潜在的治疗靶点，AMTB 可以通过抑制 TGFβ 信号通路抑制骨肉瘤细胞的生长和转移，并增加肿瘤细胞对顺铂的敏感性。