DataSheet_1_Pituitary Adenylate Cyclase Activating Polypeptide Has Inhibitory Effects on Melanoma Cell Proliferation and Migration In Vitro.pdf

Pituitary adenylate cyclase activating polypeptide (PACAP) is an endogenous neuropeptide which is distributed throughout the body. PACAP influences development of various tissues and exerts protective function during cellular stress and in some tumour formation. No evidence is available on its role in neural crest derived melanocytes and its malignant transformation into melanoma. Expression of PACAP receptors was examined in human skin samples, melanoma lesions and in a primary melanocyte cell culture. A2058 and WM35 melanoma cell lines, representing two different stages of melanoma progression, were used to investigate the effects of PACAP. PAC1 receptor was identified in melanocytes in vivo and in vitro and in melanoma cell lines as well as in melanoma lesions. PACAP administration did not alter viability but decreased proliferation of melanoma cells. With live imaging random motility, average speed, vectorial distance and maximum distance of migration of cells were reduced upon PACAP treatment. PACAP administration did not alter viability but decreased proliferation capacity of melanoma cells. On the other hand, PACAP administration decreased the migration of melanoma cell lines towards fibronectin chemoattractant in the Boyden chamber. Furthermore, the presence of the neuropeptide inhibited the invasion capability of melanoma cell lines in Matrigel chambers. In summary, we provide evidence that PACAP receptors are expressed in melanocytes and in melanoma cells. Our results also prove that various aspects of the cellular motility were inhibited by this neuropeptide. On the basis of these results, we propose PACAP signalling as a possible target in melanoma progression.

垂体腺苷酸环化酶激活多肽（Pituitary adenylate cyclase activating polypeptide，简称PACAP）是一种广泛分布于全身的固有神经肽。PACAP在多种组织的发育过程中发挥作用，并在细胞应激和某些肿瘤形成过程中发挥保护性功能。目前尚无关于其在神经嵴起源的黑色素细胞及其恶变成为黑色素瘤过程中的作用的相关证据。研究者对人类皮肤样本、黑色素瘤病变以及原代黑色素细胞培养中PACAP受体的表达进行了检测。A2058和WM35黑色素瘤细胞系，分别代表黑色素瘤进展的两个不同阶段，被用于研究PACAP的影响。在黑色素细胞、黑色素瘤细胞系以及黑色素瘤病变中均发现了PAC1受体。PACAP的给予并未改变细胞的存活率，但降低了黑色素瘤细胞的增殖能力。通过活细胞成像技术，细胞在PACAP处理后的随机运动、平均速度、向量距离和最大移动距离均有所减少。PACAP的给予并未改变细胞的存活率，但降低了黑色素瘤细胞向纤维连接蛋白趋化因子迁移的能力。此外，该神经肽的存在抑制了黑色素瘤细胞系在Matrigel培养皿中的侵袭能力。综上所述，本研究提供了PACAP受体在黑色素细胞和黑色素瘤细胞中表达的证据。我们的研究结果还证实了该神经肽抑制了细胞运动的多方面。基于这些结果，我们提出PACAP信号通路可能是黑色素瘤进展中的一个潜在治疗靶点。