薄荷醇抑制前列腺癌DU145细胞的增殖和迁移

Menthol inhibits the proliferation and motility of prostate cancer DU145 cells

目的探讨薄荷醇对雄激素非依赖性前列腺癌DU145细胞增殖和迁移能力的影响。方法通过RT-PCR、免疫组织化学和Western blot方法检测TRPM8和TRPA1的表达;MTT和划痕试验检测薄荷醇对DU145细胞的增殖和迁移能力的影响;流式细胞术检测TRPM8对DU145细胞周期和凋亡的影响。结果 RT-PCR、免疫组织化学和Western blot提示TRPM8在DU145细胞中高表达而TRPA1在DU145细胞中不表达;薄荷醇能诱导细胞周期阻滞于G0/G1期,与未经薄荷醇处理的细胞相比,经100μmol/L薄荷醇处理的细胞培养24、48和72h后,G0/G1期细胞显著增加(49.12%±1.92%vs61.71%±2.70%、77.65%±1.63%、71.81%±2.46%,P<0.05,P<0.01),进而抑制细胞增殖(P<0.05),并抑制细胞迁移(P<0.05),流式细胞术检测显示薄荷醇并不引起细胞凋亡。结论 TRPM8可能成为前列腺癌治疗的一个新靶点,对于高表达TRPM8的雄激素非依赖性前列腺癌针对TRPM8通道的药物治疗可能比TRPM8基因治疗更为实用,因此,薄荷醇作为一个潜在的抗肿瘤药物也拥有很大的发展前景。

Objective To investigate the effects of menthol on the proliferation and motility of androgen-independent prostate cancer DU145 cells. Methods The expressions of TRPM8 and TR- PAl in DU145 cells were detected using RT-PCR, immunohistoehemical assay and Western blot as say; MTT assay and scratch motility assay were performed to investigate the effects of menthol on the proliferation and motility of DU145 cells and cell cycle and cell apoptosis were also detected by flow cytometry. Results The outcome of RT-PCR, immunohistochemical assay and Western blot assay indicated that DU145 cells have a remarkable expression of TRPM8 but dose not express TRPA1. Compared with non-menthol-treated cells, there is a significant increase in G0/G1 phase cell fraction after 24, 48 and 72 h 100μmol/L menthol treatment （49.12±1.92% vs 61.71%±2. 70 %, 77.65%±1.63%, 71.81%±2.46%, P〈0.05, P〈0.01）, and menthol could inhibit the cell growth （P〈0.05） and cell motility （P〈0.05） and it failed to induce cell apoptosis. Conclusions TRPM8 may serve as important target for the treatment of prostate cancer and TRPM8 activation is more pragmatic than TRPM8 gene therapy for those androgen-independent prostate cancers with high level expression of TRPM8. Thus, menthol is a useful compound for future development as an anticancer agent.