Research Laboratory on Neuropeptides, Virgen del Rocío University Hospital, Sevilla, Spain.
Lab Invest. 2010 Aug;90(8):1259-69. doi: 10.1038/labinvest.2010.92. Epub 2010 May 10.
Melanoma, the most deadly form of skin cancer, is aggressive and resistant to current therapies. It has been previously reported that the substance P and neurokinin-1 (NK-1) receptor antagonists induce cell proliferation and cell inhibition, respectively, in human melanoma cell lines. Aprepitant is a selective high-affinity antagonist of the human NK-1 receptor. Until now, this drug has been used as an anxiolytic, antidepressant and antiemetic. Moreover, the antitumor action of aprepitant has been previously reported. However, the presence of NK-1 receptors in human melanomas and whether the antitumor action of the NK-1 receptor antagonist aprepitant is exerted on human malignant melanomas have not been previously described. The aims of this study are to show the presence of NK-1 receptors in human malignant melanomas and the antitumoral action of aprepitant against several human melanoma cell lines. Immunoblot analysis was used to determine the presence of NK-1 receptors in human melanoma cell lines, and immunohistochemistry was used to demonstrate NK-1 receptors in human melanoma samples. We performed an in vitro study of the cytotoxicity of the NK-1 receptor antagonist aprepitant on human melanoma cell lines. A coulter counter was used to determine viable cell numbers, followed by application of the tetrazolium compound MTS. The DAPI method was applied to demonstrate apoptosis. We observed that NK-1 receptors were present in all the melanoma samples studied as well as in human melanoma cell lines. We also showed that melanoma cell lines expressed mRNA for the NK-1 receptor. Moreover, after using a knockdown method, we showed that NK-1 receptors are involved in the viability of tumor cells. In this study, we also report that aprepitant, at 10-60 microM concentrations, elicits cell growth inhibition in a concentration-dependent manner in all melanoma cell lines studied, that the specific antitumor action of aprepitant occurs through the NK-1 receptor and that melanoma cell death is due to apoptosis. These findings show for the first time that the NK-1 receptor may be a promising new target and that the NK-1 receptor antagonist aprepitant could be a candidate as a new antitumor drug in the treatment of human melanoma.
黑色素瘤是最致命的皮肤癌形式，具有侵袭性且对当前的治疗方法具有抗性。先前有报道称，P 物质和神经激肽-1（NK-1）受体拮抗剂分别诱导人黑色素瘤细胞系的细胞增殖和细胞抑制。阿瑞匹坦是一种选择性高亲和力的人 NK-1 受体拮抗剂。到目前为止，这种药物已被用作抗焦虑药、抗抑郁药和止吐药。此外，先前已经报道了阿瑞匹坦的抗肿瘤作用。然而，NK-1 受体在人黑色素瘤中的存在以及 NK-1 受体拮抗剂阿瑞匹坦的抗肿瘤作用是否对人恶性黑色素瘤发挥作用尚未被描述。本研究的目的是证明 NK-1 受体存在于人恶性黑色素瘤中，以及阿瑞匹坦对几种人黑色素瘤细胞系的抗肿瘤作用。免疫印迹分析用于确定人黑色素瘤细胞系中 NK-1 受体的存在，免疫组织化学用于证明人黑色素瘤样本中的 NK-1 受体。我们对 NK-1 受体拮抗剂阿瑞匹坦对人黑色素瘤细胞系的细胞毒性进行了体外研究。库尔特计数器用于确定活细胞数量，然后应用四唑化合物 MTS。应用 DAPI 法证明细胞凋亡。我们观察到 NK-1 受体存在于所有研究的黑色素瘤样本以及人黑色素瘤细胞系中。我们还表明，黑色素瘤细胞系表达 NK-1 受体的 mRNA。此外，在用敲低方法后，我们表明 NK-1 受体参与肿瘤细胞的活力。在这项研究中，我们还报告说，阿瑞匹坦在 10-60μM 浓度下以浓度依赖性方式在所有研究的黑色素瘤细胞系中引起细胞生长抑制，阿瑞匹坦的特异性抗肿瘤作用通过 NK-1 受体发生，并且黑色素瘤细胞死亡是由于细胞凋亡。这些发现首次表明 NK-1 受体可能是一个有前途的新靶点，NK-1 受体拮抗剂阿瑞匹坦可能成为治疗人类黑色素瘤的新型抗肿瘤药物的候选药物。
