吡咯烷二硫代氨基甲酸盐增强三氧化二砷对急性早幼粒细胞白血病细胞的细胞毒性作用。
Pyrrolidine Dithiocarbamate Enhances the Cytotoxic Effect of Arsenic Trioxide on Acute Promyelocytic Leukemia Cells.
机构信息
Department of Traditional Chinese Medicine, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Department of Cardiology, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
出版信息
Comb Chem High Throughput Screen. 2023;26(11):2067-2076. doi: 10.2174/1386207326666230123155944.
BACKGROUND
More than 95% patients with acute promyelocytic leukemia (APL) carry the PML-RARα fusion oncoprotein. Arsenic trioxide (ATO) is an efficacious therapeutic agent for APL, and the mechanism involves the binding with PML and degradation of PML-RARα protein. Pyrrolidine dithiocarbamate (PDTC) demonstrates the function of facilitating the cytotoxic effect of ATO.
PURPOSE
To investigate whether PDTC is potential to enhance the cytotoxic effect of ATO to APL cells by acting on PML-RARα oncoproteins.
METHODS
Inhibitory effects of drugs on cell viability were examined by CCK-8 test, and apoptosis was evaluated by flow cytometry. Western blotting and immunofluorescence assays were used to explore the mechanism.
RESULTS
PDTC improved the effect of ATO on inhibiting proliferation of NB4 cells in vitro. Further, PDTC-ATO promoted apoptosis and cell cycle arrest in NB4 cells. The expression of caspase- 3 and Bcl-2 was reduced in PDTC-ATO-treated NB4 cells, while cleaved caspase-3 and Bax was up-regulated. Furthermore, less PML-RARα expression were found in PDTC-ATO-treated NB4 cells than that in NB4 cells treated with ATO singly. Laser confocal microscopy showed that protein colocalization of PML and RARα was disrupted more significantly by PDTC-ATO treatment than that with ATO monotherapy.
CONCLUSION
In conclusion, PDTC enhanced the cytotoxic effect of ATO on APL, and the mechanism was, at least in part, related to the promotion of ATO-induced degradation of PML-RARα fusion protein via forming a complex PDTC-ATO.
背景
超过 95%的急性早幼粒细胞白血病（APL）患者携带 PML-RARα 融合癌蛋白。三氧化二砷（ATO）是治疗 APL 的有效治疗药物，其机制涉及与 PML 结合和降解 PML-RARα 蛋白。吡咯烷二硫代氨基甲酸盐（PDTC）表现出促进 ATO 细胞毒性作用的功能。
目的
研究 PDTC 是否通过作用于 PML-RARα 癌蛋白来增强 ATO 对 APL 细胞的细胞毒性作用。
方法
通过 CCK-8 试验检测药物对细胞活力的抑制作用，通过流式细胞术评估细胞凋亡。Western blot 和免疫荧光分析用于探索机制。
结果
PDTC 改善了 ATO 在体外抑制 NB4 细胞增殖的作用。此外，PDTC-ATO 促进 NB4 细胞凋亡和细胞周期停滞。PDTC-ATO 处理的 NB4 细胞中 caspase-3 和 Bcl-2 的表达减少，而 cleaved caspase-3 和 Bax 上调。此外，与单独用 ATO 处理的 NB4 细胞相比，PDTC-ATO 处理的 NB4 细胞中 PML-RARα 的表达减少。激光共聚焦显微镜显示，与单独用 ATO 治疗相比，PDTC-ATO 处理更显著地破坏了 PML 和 RARα 蛋白的共定位。
结论
总之，PDTC 增强了 ATO 对 APL 的细胞毒性作用，其机制至少部分与通过形成 PDTC-ATO 复合物促进 ATO 诱导的 PML-RARα 融合蛋白降解有关。
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