Shannan B, Seifert M, Boothman D A, Tilgen W, Reichrath J
Department of Dermatology, The Saarland University Hospital, Building 18, D-66421 Homburg, Germany.
J Steroid Biochem Mol Biol. 2007 Mar;103(3-5):721-5. doi: 10.1016/j.jsbmb.2006.12.068. Epub 2007 Jan 16.
Prostate cancer is the most commonly diagnosed cancer in the majority of western countries. Due to their antiproliferative and proapoptotic activity, vitamin D analogues have been introduced recently as an experimental therapy for prostate cancer. Clusterin (CLU) is a glycoprotein that has two known isoforms generated in human cells. A nuclear form of CLU protein (nCLU) is pro-apoptotic, and a secretory form (sCLU) is pro-survival. In this study, we analyzed whether proapoptotic and antiproliferative effects of 1,25(OH)(2)D(3) on LNCaP prostate cancer cells are modulated by expression of sCLU. Using colony forming assay, we studied the effect of treatment with different doses of 1,25(OH)(2)D(3) (10(-6), 10(-7), 10(-10)M) on proliferation of LNCaP cells that were stable transfected and over-express sCLU (LNT-1) as compared to empty vector-transfected cells (LN/C). We also measured apoptosis using TUNEL assay. sCLU over-expression protected against both antiproliferative (30%) and proapoptotic (15%) effects of 1,25(OH)(2)D(3), although this effect was statistically not significant. In conclusion, our findings demonstrate that expression of sCLU modulates growth regulatory effects of 1,25(OH)(2)D(3) in prostate cancer indicating that CLU interferes with vitamin D signalling pathways.
在大多数西方国家,前列腺癌是最常被诊断出的癌症。由于具有抗增殖和促凋亡活性,维生素D类似物最近已被引入作为前列腺癌的一种实验性疗法。聚集素(CLU)是一种糖蛋白,在人类细胞中有两种已知的异构体。CLU蛋白的核形式(nCLU)具有促凋亡作用,而分泌形式(sCLU)具有促生存作用。在本研究中,我们分析了1,25(OH)₂D₃对LNCaP前列腺癌细胞的促凋亡和抗增殖作用是否受sCLU表达的调节。我们使用集落形成试验,研究了用不同剂量的1,25(OH)₂D₃(10⁻⁶、10⁻⁷、10⁻¹⁰M)处理对稳定转染并过表达sCLU的LNCaP细胞(LNT-1)增殖的影响,并与空载体转染细胞(LN/C)进行比较。我们还使用TUNEL试验检测了细胞凋亡情况。sCLU的过表达对1,25(OH)₂D₃的抗增殖作用(30%)和促凋亡作用(15%)均有保护作用,尽管这种作用在统计学上不显著。总之,我们的研究结果表明,sCLU的表达调节了1,25(OH)₂D₃在前列腺癌中的生长调节作用,这表明CLU干扰了维生素D信号通路。