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目的探讨三氧化二砷(As203,ATO)联合Cyclopamine(CYA)对非雄激素依赖前列腺癌PC3细胞周期的影响及其可能的机制。方法不同浓度ATO、CYA单用及联合处理细胞24~72h,采用MTT比色法检测细胞增殖活性;流式细胞仪检测细胞周期,Real-time PCR及免疫荧光检测PC3细胞内Hedgehog(Hh)信号通路中相关蛋白Smoothened(Smo)、Patched(Ptch)及GLI1的表达情况。结果 MTT结果显示ATO联合CYA协同抑制PC3细胞生长,流式细胞术检测发现CYA可明显增强ATO引起PC3细胞S期阻滞作用;Real-time PCR及免疫荧光分析结果显示ATO联合CYA较单剂量组明显下调Hh信号通路中相关蛋白的表达。结论 CYA联合ATO可协同抑制PC3细胞生长,影响细胞周期进展,将其阻滞于S期,其作用机制可能是两药联合协同下调Hh信号通路活性。
Objective To investigate the effect of arsenic trioxide (As2O3, ATO) combined with Cyclopamine (CYA) on the cell cycle of androgen-independent prostate cancer PC3 cells and its possible mechanism. Methods Cells were treated with ATO and CYA at different concentrations for 24-72 h. MTT assay was used to detect cell proliferation. Cell cycle was detected by flow cytometry. Hedgehog (Hh) signal was detected by Real-time PCR and immunofluorescence staining Pathway related proteins Smoothened (Smo), Patched (Ptch) and GLI1 expression. Results MTT assay showed that ATO combined with CYA inhibited the growth of PC3 cells. Flow cytometry showed that CYA significantly increased the S phase arrest of PC3 cells induced by ATO. Real-time PCR and immunofluorescence analysis showed that ATO combined with CYA Obviously down-regulated the expression of related proteins in Hh signaling pathway. Conclusions CYA combined with ATO can inhibit the growth of PC3 cells and affect the cell cycle progression, arresting it in S phase. The mechanism may be that the combination of CYA and ATO can down-regulate the activity of Hh signaling pathway.