Sanchez Maria G, Sanchez Ana M, Collado Beatriz, Malagarie-Cazenave Sophie, Olea Nuria, Carmena Maria J, Prieto Juan C, Diaz-Laviada I Ines
Department of Biochemistry and Molecular Biology, School of Medicine, University of Alcala, Alcala de Henares, 28871 Madrid, Spain.
Eur J Pharmacol. 2005 May 16;515(1-3):20-7. doi: 10.1016/j.ejphar.2005.04.010.
Vanilloid receptor subtype-1 (TRPV1), the founding member of the vanilloid receptor-like transient receptor potential channel family, is a non-selective cation channel that responds to noxious stimuli such as low pH, painful heat and irritants. In the present study, we show, as means of reverse transcriptase-polymerase chain reaction and Western blot analysis, that the vanilloid TRPV1 receptor is expressed in the prostate epithelial cell lines PC-3 and LNCaP as well as in human prostate tissue. The kinetic parameters inferred from [(125)I]-resiniferatoxin binding were in concordance with data of TRPV1 receptors expressed in other tissues. The contribution of the endogenously expressed TRPV1 channel to intracellular calcium concentration increase in the prostate cells was studied by measuring changes in Fura-2 fluorescence by fluorescence microscopy. Addition of capsaicin, (R)-methanandamide and resiniferatoxin to prostate cells induced a dose-dependent increase in the intracellular calcium concentration that was reversed by the vanilloid TRPV1 receptor antagonist capsazepine. These results indicate that the vanilloid TRPV1 receptor is expressed and functionally active in human prostate cells.
香草酸受体亚型1(TRPV1)是香草酸受体样瞬时受体电位通道家族的首个成员,是一种对诸如低pH值、灼痛热和刺激物等有害刺激产生反应的非选择性阳离子通道。在本研究中,我们通过逆转录聚合酶链反应和蛋白质免疫印迹分析表明,香草酸TRPV1受体在前列腺上皮细胞系PC-3和LNCaP以及人类前列腺组织中表达。从[(125)I] - 树脂毒素结合推断出的动力学参数与在其他组织中表达的TRPV1受体的数据一致。通过荧光显微镜测量Fura-2荧光的变化,研究了内源性表达的TRPV1通道对前列腺细胞内钙浓度增加的作用。向前列腺细胞中添加辣椒素、(R)-甲烷酰胺和树脂毒素会导致细胞内钙浓度呈剂量依赖性增加,而香草酸TRPV1受体拮抗剂辣椒平可逆转这种增加。这些结果表明,香草酸TRPV1受体在人类前列腺细胞中表达且具有功能活性。