补骨脂酚拮抗AR转录活性抑制雄激素诱导的前列腺癌细胞LNCaP的增殖

Bakuchiol inhibits the androgen induced-proliferation of prostate cancer cell line LNCaP through suppression of AR transcription activity

关键词:  补骨脂酚  前列腺癌  补骨脂  雄激素受体拮抗剂

Key words:  Bakuchiol  prostate cancer  Psoralea corylifolia L.  antagonist of androgen receptor

摘要:

[目的] 研究补骨脂酚对雄激素受体(AR)的亲和性和转录活性的调控在雄激素依赖的前列腺癌细胞LNCaP中的作用。[方法] 用AR竞争性结合试剂盒检测补骨脂酚结合AR的能力;用荧光素酶报告质粒法检测补骨脂酚对睾酮诱导的AR转录活性的影响;用实时定量PCR法检测补骨脂酚对LNCaP细胞中雄激素应答的靶基因——前列腺特异抗原(PSA)表达的影响;用MTT法检测补骨脂酚对睾酮诱导的LNCaP细胞增殖的影响。[结果] 补骨脂酚体外结合AR的能力与AR拮抗剂氟他胺相当;睾酮诱导的AR转录活性可以被补骨脂酚阻断;补骨脂酚可以抑制睾酮对LNCaP细胞PSA表达和增殖的上调作用。[结论] 补骨脂酚经由AR抑制雄激素依赖的LNCaP细胞的增殖和基因表达,提示:补骨脂酚可以作为潜在的AR拮抗剂,用于雄激素依赖的PCa药物的开发。

Abstract:

[Objective] To explore the effect of bakuchiol on the affinity and transcription activity of androgen receptor (AR), and by which bakuchiol behaviors in the androgen-dependent PCa cell line LNCaP. [Methods] AR competitor binding assay was used to detect the affinity of bakuchiol to AR. Luciferase reporter assay was used to detect the effect of bakuchiol on testosterone-induced AR transcription activity. LNCaP cells were treated with bakuchiol and/or testosterone and the expression of androgen targeted gene prostate specific antigen (PSA) and the cell proliferation induced by testosterone were then detemined by real-time PCR and MTT assay. [Results] The IC₅₀ of bakuchiol to AR is 8.87×10⁴, which is similar with flutamide (10.00×10⁴). Bakuchiol effectively blocked testosterone-induced AR transcription activity. The PSA expression and cell proliferation induced by testosterone in LNCaP were significantly inhibited by bakuchiol. [Conclusion] Bakuchiol suppressed testosterone-induced cell proliferation and gene expression in LNCaP by targeting AR, which indicates a potential usefulness of bakuchiol as an AR antagonist for drug development and treatment of androgen-dependent PCa.