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目的:探讨TRPM8抑制剂BCTC对前列腺癌DU145细胞的抑癌作用。方法:PCR和Western blot检测TRPM8的表达;MTT法检测DU145细胞增殖能力;划痕实验、transwell实验检测细胞迁移和侵袭能力;流式细胞术检测细胞周期和凋亡;Western blot检测周期相关蛋白(p-AKT,p-GSK-3β,cyclin B1,cyclin D1,CDK2/4/6)、凋亡相关蛋白(Bcl-2,Bax,caspase-3)、迁移相关蛋白(pFAK,MMP2)表达水平的变化。结果:PCR和Western blot提示TRPM8在前列腺癌DU145细胞中表达显著高于正常前列腺上皮PNT1A细胞;BCTC处理后,DU145细胞增殖能力下降(P<0.05),细胞周期表现为G0/G1阻滞期(P<0.05),迁移、侵袭能力也明显下降(P<0.05),但凋亡相关实验显示BCTC并不引起细胞凋亡(P>0.05)。Western blot显示BCTC能下调p-AKT,cyclin D1,CDK2,CDK6,MMP2和pFAK等的表达,而上调p-GSK-3β的表达。结论:TRPM8抑制剂BCTC能抑制前列腺癌DU145细胞增殖、迁移和侵袭能力,是一个极具潜力的抗前列腺癌药物。
Objective: To investigate the tumor suppressor effect of TRPM8 inhibitor BCTC on prostate cancer DU145 cells. METHODS: The expression of TRPM8 was detected by PCR and Western blot. The proliferation of DU145 cells was detected by MTT assay. The scratch and transwell assays were used to detect the cell migration and invasion. Cell cycle and apoptosis were detected by flow cytometry. The expressions of p-AKT, p-GSK-3β, cyclin B1, cyclin D1, CDK2 / 4/6 and Bcl-2, Bax and caspase- Variety. Results: PCR and Western blot suggested that TRPM8 expression in prostate cancer DU145 cells was significantly higher than that in normal prostate epithelium PNT1A cells. After BCTC treatment, the proliferation of DU145 cells decreased (P <0.05), and the cell cycle showed G0 / G1 arrest P <0.05), migration and invasion ability also significantly decreased (P <0.05), but apoptosis related experiments showed that BCTC did not cause apoptosis (P> 0.05). Western blot showed that BCTC down-regulated the expression of p-AKT, cyclin D1, CDK2, CDK6, MMP2 and pFAK, and up-regulated the expression of p-GSK-3β. Conclusion: The TRPM8 inhibitor BCTC can inhibit the proliferation, migration and invasion of prostate cancer DU145 cells. It is a potential anti-prostate cancer drug.