■据报道,低甲基化剂(HMA)靶向骨髓增生异常综合征(MDS)中的SonicHedgehog(Shh)信号通路。然而,在MUTZ-1细胞系中,Smo抑制剂Jervine及其与地西他滨的组合仍然缺乏协同抑制作用。
■我们使用CCK-8测定法来检测MUTZ-1细胞系的体外增殖率。此外,采用AnnexinV-FITC/PI双染色流式细胞仪检测细胞凋亡率和细胞周期变化。用qRT-PCR定量mRNA的表达水平,蛋白质印迹法检测蛋白质的表达。
■我们发现单剂Jervine或地西他滨可以显着抑制MUTZ-1细胞系的增殖率,这种抑制作用是时间依赖性和浓度依赖性的。沙文和地西他滨联合干预能更显著地抑制细胞增殖,诱导细胞凋亡,并阻断细胞周期的G1期。两种药物的联合干预可显著降低MUTZ-1细胞中Smo和G1i-1mRNA的表达。此外,在结合两种药物治疗后,Smo的蛋白质水平,G1i-1,PI3K,p-AKT,Bcl2和细胞周期蛋白Dl显著下调,Caspase-3上调,这表明泽文与地西他滨的组合可能对控制增殖有效,凋亡,和细胞周期。
■Smo抑制剂Jervine及其与地西他滨的组合对增殖具有协同作用,细胞周期,MUTZ-1细胞凋亡,其机制可能通过干扰Shh信号通路来实现。
UNASSIGNED: Hypomethylating agents (HMAs) have been reported to target the Sonic Hedgehog (Shh) signaling pathway in myelodysplastic syndrome (MDS). However, the synergistic inhibitory effect of Smo inhibitor jervine and its combination with decitabine in MUTZ-1 cell lines remains lacking.
UNASSIGNED: We used a CCK-8 assay to detect the in-vitro proliferation rate of MUTZ-1 cell lines. Besides, the Annexin V-FITC/PI double staining flow cytometry was utilized to detect the apoptosis rate and cell cycle changes. The expression levels of mRNA were quantified by using qRT-PCR, and the western blot was employed to detect the expression of proteins.
UNASSIGNED: We found that the single-agent jervine or decitabine can significantly inhibit the proliferation rate of MUTZ-1 cell lines, and this inhibitory effect is time-dependent and concentration-dependent. The combined intervention of the jervine and decitabine can more significantly inhibit cell proliferation, induce cell apoptosis, and block the G1 phase of the cell cycle. The combined intervention of the two drugs significantly reduced Smo and G1i-1 mRNA expression in MUTZ-1 cells. Furthermore, after combining both of the drug treatments, the proteins levels of Smo, G1i-1, PI3K, p-AKT, Bcl2, and Cyclin Dl were significantly downregulated, and Caspase-3 is upregulated, indicating that jervine with its combination of decitabine might be effective for controlling the proliferation, apoptosis, and cell cycle.
UNASSIGNED: The Smo inhibitor jervine and its combination with decitabine have a synergistic effect on the proliferation, cell cycle, and apoptosis of MUTZ-1 cells, and its mechanism may be achieved by interfering with the Shh signaling pathway.