Department of Clinical Sciences, Division of Urologic Research, Malmö University Hospital, Lund University, Malmö, Sweden.
Urol Oncol. 2011 Jul-Aug;29(4):436-45. doi: 10.1016/j.urolonc.2009.09.013. Epub 2009 Nov 19.
This study was conducted to examine the effects of 5-HT on extracellular signal-regulated kinase 1/2 (Erk1/2) and Akt pathways in prostate cancer (PC) cells.
PC cell lines PC-3, Du145, and LNCaP stimulated with 5-HT in the presence of MEK or PI3K inhibitors and 5-HT receptor subtype 1A antagonist were analyzed by Western blotting and immunofluorescence. The proliferation assay BrdU and Boyden chamber were used to determine proliferation and migration, respectively.
5-HT dose-dependently induced rapid activation of Erk1/2 in PC-3 and Du145 cells, whereas in LNCaP cells, Erk1/2 phosphorylation was slow and sustained for up to 18 h. Similarly, 5-HT induced phosphorylation of Akt within 1 hour of stimulation, however, Akt phosphorylation was more pronounced in Du145 cells compared with PC-3 or LNCaP cells. The action of 5-HT was inhibited to varying degrees by inhibitors of MAPK and PI3K as well as by a 5-HT receptor subtype 1A antagonist. In addition to proliferation, 5-HT induced migration of PC-3 and Du145 cells, which were alleviated by the aforementioned inhibitors. The effects of 5-HT on LNCaP cells appeared to be related to neuroendocrine-phenotype acquisition and chromogranin A and neuron specific enolase expression.
This study addresses the role of 5-HT in Erk1/2 and Akt activation in PC cells. The data presented here identify 5-HT receptors as a novel target in castration-resistant PC. Furthermore, our observations are in line with previous studies, which point towards neuroendocrine factors facilitating progression and migration of prostatic cancer cells in an androgen-deficient environment. Nonetheless, additional studies are warranted to corroborate the role of 5-HTR antagonists as a potential target for anticancer therapy.
本研究旨在探讨 5-羟色胺（5-HT）对前列腺癌细胞外信号调节激酶 1/2（Erk1/2）和 Akt 通路的影响。
采用 Western blot 和免疫荧光法分析经 5-HT 刺激的 PC-3、Du145 和 LNCaP 细胞系，同时加入 MEK 或 PI3K 抑制剂和 5-HT 受体亚型 1A 拮抗剂。BrdU 增殖实验和 Boyden 室迁移实验分别用于测定细胞增殖和迁移。
5-HT 呈剂量依赖性诱导 PC-3 和 Du145 细胞中 Erk1/2 的快速激活，而在 LNCaP 细胞中，Erk1/2 的磷酸化则缓慢且可持续长达 18 小时。同样，5-HT 在刺激后 1 小时内诱导 Akt 的磷酸化，但与 PC-3 或 LNCaP 细胞相比，Du145 细胞中 Akt 的磷酸化更为明显。MAPK 和 PI3K 抑制剂以及 5-HT 受体亚型 1A 拮抗剂均可不同程度地抑制 5-HT 的作用。除增殖外，5-HT 还诱导 PC-3 和 Du145 细胞迁移，上述抑制剂可减轻其迁移作用。5-HT 对 LNCaP 细胞的作用似乎与获得神经内分泌表型以及嗜铬粒蛋白 A 和神经元特异性烯醇化酶的表达有关。
本研究探讨了 5-HT 在 PC 细胞中 Erk1/2 和 Akt 激活中的作用。本研究鉴定出 5-HT 受体是去势抵抗性前列腺癌的一个新靶点。此外，我们的观察结果与先前的研究一致，这些研究表明神经内分泌因子在雄激素缺乏环境中促进前列腺癌细胞的进展和迁移。然而，还需要进一步的研究来证实 5-HT 受体拮抗剂作为一种潜在的抗癌治疗靶点的作用。
