The role of serotonin (5-hydroxytryptamine1A and 1B) receptors in prostate cancer cell proliferation
The role of serotonin (5-hydroxytryptamine1A and 1B) receptors in prostate cancer cell proliferation
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DOI：
10.1016/j.juro.2006.06.087
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发表时间：
2006-10-01
影响因子：
6.6
通讯作者：
Mumtaz, Faiz H.
中科院分区：
文献类型：
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作者：
Siddiqui, Emad J.;Shabbir, Majid;Mumtaz, Faiz H.
目的：5-羟色胺（5-hydroxytryptamine， 5-羟色胺）是前列腺神经内分泌细胞释放的单胺类神经递质，在肿瘤的生长、分化和基因表达中具有重要作用。我们研究了5-羟色胺和5-羟色胺拮抗剂对前列腺癌细胞生长的影响，并在PC3细胞和人激素难治性前列腺癌组织中鉴定了5-羟色胺受体的表达。材料与方法：用5-羟色胺或5-羟色胺受体拮抗剂5-羟色胺（1A）、（1B）、（1D）、(2)、(3)、(4)孵育12种激素难治性PC3人前列腺癌细胞。72h后用结晶紫法测定细胞活力。流式细胞术观察5-羟色胺（1A）和（1B）拮抗剂处理的PC3细胞凋亡情况。采用免疫组织化学和Western blot方法对激素难治性人前列腺癌组织PC3细胞及其切片进行了研究。结果：在PC3细胞中，浓度为10(-8)M的12种制剂中，5-羟色胺可引起剂量依赖性增殖，72小时时，与对照组相比，5-羟色胺最大增加15% （p < 0.0001）。5HT（1A）拮抗剂na -190氢溴化物和5-羟色胺（1B）拮抗剂SB224289 HCl （Tocris实验室，Bristol，英国）在浓度为10(-4)M时，72h对PC3细胞生长的抑制作用分别比对照组高20%和78% （p < 0.0001）。在PC3细胞中，5-羟色胺（1A）和（1B）拮抗剂在孵卵24小时和48小时后显示凋亡。在PC3细胞和前列腺癌组织中可见5-羟色胺（1A）和（1B）受体的免疫染色。Western blot分析显示，5-羟色胺，A和，受体蛋白分别为46和43 kDa带。结论：在PC3前列腺癌细胞中，5-羟色胺（1A）和更大程度上5-羟色胺（1B）拮抗剂显著抑制生长并诱导凋亡。据我们所知，5-羟色胺（1B）拮抗剂SB224289 HCl引起的生长抑制是一个新发现，2种拮抗剂5-羟色胺（1A）和（1B）引起的细胞凋亡也是一个新发现。这种作用很可能是通过5-羟色胺（1A）和受体介导的。因此，我们的研究结果表明5-羟色胺（1A），特别是5-羟色胺（1B）受体拮抗剂作为潜在的抗肿瘤药物值得进一步研究。
Purpose: Serotonin (5-hydroxytryptamine), a monoamine neurotransmitter released by prostate neuroendocrine cells, has a fundamental role in tumor growth, differentiation and gene expression. We investigated the effect of 5-hydroxytryptamine and 5-hydroxytryptamine antagonists on the growth of prostate cancer cells and we identified 5-hydroxytryptamine receptor expression in PC3 cells and in human hormone refractory prostate cancer tissue.Materials and Methods: A total of 12 preparations of hormone refractory PC3 human prostate cancer cells were incubated with 5-hydroxytryptamine, or the 5-hydroxytryptamine receptor antagonists 5-hydroxytryptamine(1A), (1B), (1D), (2), (3) or (4). After 72 hours cell viability was assessed using the crystal violet assay. PC3 cells treated with 5-hydroxytryptamine(1A) and (1B) antagonists were investigated for apoptosis using flow cytometry. PC3 cells and sections of hormone refractory human prostate cancer tissue were studied by immunohistochemistry and Western blot analysis.Results: In PC3 cells 5-hydroxytryptamine caused dose dependent proliferation with a maximum increase of 15% in 12 preparations at a concentration of 10(-8) M at 72 hours compared to controls (p < 0.0001). At a concentration of 10(-4) M at 72 hours the 5HT(1A) antagonist NAN-190 hydrobromide and the 5-hydroxytryptamine(1B) antagonist SB224289 HCl (Tocris Laboratories, Bristol, United Kingdom) induced a 20% and 78% inhibitory effect, respectively, on PC3 cell growth compared to that in controls (p < 0.0001). In PC3 cells 5-hydroxytryptanline(1A) and (1B) antagonists demonstrated apoptosis after 24 and 48 hours of incubation. Immunostaining for 5-hydroxytryptamine(1A) and (1B) receptors was seen in PC3 cells and prostate cancer tissue. Western blot analysis demonstrated 5-hydroxytryptamine,A and, receptor proteins with 46 and 43 kDa bands, respectively.Conclusions: In PC3 prostate cancer cells 5-hydroxytryptamine(1A) and to a greater extent 5-hydroxytryptamine(1B) antagonists significantly inhibit growth and induce apoptosis. To our knowledge growth inhibition caused by the 5-hydroxytryptamine(1B) antagonist SB224289 HCl is a novel finding, as is apoptosis caused by the 2 antagonists 5-hydroxytryptamine(1A) and (1B). This effect is most likely mediated via 5-hydroxytryptamine(1A) and, receptors. Therefore, our results imply that 5-hydroxytryptamine(1A) and in particular 5-hydroxytryptamine(1B) receptor antagonists warrant further investigations as potential anti-neoplastic agents.