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目的明确去甲斑蝥素(norcantharidin,NCTD)对肿瘤细胞周期的影响,进一步阐明NCTD的抗肿瘤作用机制。方法体外培养HL-60细胞,NCTD作用后采用3H-TdR掺入实验检测DNA复制;流式细胞术检测细胞周期进程;Western blotting检测细胞周期调控蛋白Cdk1、Cyclin B1、Cdc 25c表达变化。结果不同浓度(16~128μmol/L)NCTD作用于HL-60细胞12 h后,DNA复制受到抑制,细胞周期出现明显的S期累积,并呈剂量依赖性趋势;同时观察到G2/M的阻滞,且调控G2/M周期进程的调节蛋白Cdk1、Cyclin B1及Cdc 25C蛋白表达下调。结论 NCTD可抑制HL-60细胞DNA复制、下调周期调控蛋白从而干扰肿瘤细胞的周期进程。
Objective To determine the effect of norcantharidin (NCTD) on tumor cell cycle and further elucidate the antitumor mechanism of NCTD. Methods HL-60 cells were cultured in vitro. DNA replication was detected by 3H-TdR incorporation assay after NCTD treatment. Cell cycle progression was detected by flow cytometry. The expression of cell cycle regulatory proteins Cdk1, Cyclin B1 and Cdc 25c were detected by Western blotting. Results After treated with NCTD of different concentrations (16 ~ 128μmol / L) for 12 h, the DNA replication was inhibited and the cell cycle appeared obvious S phase accumulation in a dose-dependent manner. At the same time, the G2 / M resistance The expressions of Cdk1, Cyclin B1 and Cdc 25C, which regulate the G2 / M cycle progression, were down-regulated. Conclusions NCTD can inhibit the DNA replication of HL-60 cells and down-regulate the regulatory proteins to interfere with the tumor cell cycle progression.