Kim Hyun-Jung, Park Young In, Dong Mi-Sook
School of Life Sciences and Biotechnology, Korea University, 1, 5-Ka, Anam-dong, Sungbuk-ku, Seoul 136-701, Republic of Korea.
Toxicol In Vitro. 2006 Oct;20(7):1159-67. doi: 10.1016/j.tiv.2006.03.003. Epub 2006 Mar 8.
In order to select a better prostate cancer cell model for androgen receptor (AR)-mediated reporter gene assays, we assessed the androgen response characteristics of three cell lines, LNCaP, PC3/AR(+) and 22Rv1, in this study. Both the mRNA and the proteins of AR and glucocorticoid receptor (GR) were expressed in all three cell lines. Among the three cell lines, only in LNCaP cells, DHT concentration-dependently stimulated proliferation. DHT induced the luciferase activity in three cell lines which were transiently transfected with pMMTV-Luc, in a concentration-dependent manner. The maximum induction was 24.0-fold and 13.4-fold in 22Rv1 and in the LNCaP respectively. PC3/AR(+) were more sensitive to respond to DHT at a minimal concentration of 10(-12)M by 14.0-fold induction. The transcriptional activity induced with 10(-8)M DHT was inhibited about 50-75% in the PC3/AR(+) and 22Rv1, and 98% in the LNCaP, by vinclozolin. Dexamethasone concentration-dependently induced the luciferase activity in PC3 and 22Rv1, but not in the LNCaP. However, the response to dexamethasone in 22Rv1 was very weak compared to DHT. The (anti)androgencity of seven pyrethroids was assessed via an AR-mediated luciferase reporter assay. None of them showed the androgenic action in all three cell lines. Permethrin inhibited the DHT induced luciferase activity about 22%, 35.8% and 75.5% in 22Rv1, PC3/AR(+) and LNCaP, respectively. Based on results from in this study and cell line character, 22Rv1 cells seemed to be an appropriate model for the screening of androgenic endocrine disruptors, although it needs further studies with other steroid receptor and thyroid receptor.
为了选择一个更好的用于雄激素受体(AR)介导的报告基因检测的前列腺癌细胞模型,我们在本研究中评估了三种细胞系LNCaP、PC3/AR(+)和22Rv1的雄激素反应特性。AR和糖皮质激素受体(GR)的mRNA及蛋白在所有三种细胞系中均有表达。在这三种细胞系中,仅LNCaP细胞中,双氢睾酮(DHT)浓度依赖性地刺激细胞增殖。DHT以浓度依赖性方式诱导瞬时转染pMMTV-Luc的三种细胞系中的荧光素酶活性。在22Rv1和LNCaP中,最大诱导倍数分别为24.0倍和13.4倍。PC3/AR(+)在最低浓度10(-12)M的DHT作用下更敏感,诱导倍数为14.0倍。在PC3/AR(+)和22Rv1中,10(-8)M DHT诱导的转录活性被乙烯菌核利抑制约50 - 75%,在LNCaP中被抑制98%。地塞米松浓度依赖性地诱导PC3和22Rv1中的荧光素酶活性,但在LNCaP中无此作用。然而,与DHT相比,22Rv1对地塞米松的反应非常弱。通过AR介导的荧光素酶报告基因检测评估了七种拟除虫菊酯的(抗)雄激素活性。它们在所有三种细胞系中均未显示出雄激素作用。氯菊酯分别在22Rv1、PC3/AR(+)和LNCaP中抑制DHT诱导的荧光素酶活性约22%、35.8%和75.5%。基于本研究结果和细胞系特征,22Rv1细胞似乎是筛选雄激素内分泌干扰物的合适模型,尽管还需要对其他类固醇受体和甲状腺受体进行进一步研究。