[关键词]
[摘要]
目的 探讨双吲哚马来酰亚胺衍生物L6诱导白血病细胞的凋亡作用及其分子机制。方法 采用MTT比色法检测L6对白血病细胞HEL、K562、KG1a的杀伤作用。流式细胞术检测L6对HEL细胞凋亡、周期和分化的影响。Western blotting法检测细胞凋亡相关蛋白的表达。通过体内实验研究L6治疗小鼠白血病的作用效果。结果 MTT比色法结果显示L6对HEL、K562、KG1a细胞具有比阳性对照米哚妥林(PKC412)更显著的抑制活性,半数抑制浓度(IC50)分别为(0.05±0.03)、(0.32±0.01)、(0.19±0.10)μmol/L。L6可以诱导HEL细胞发生凋亡和G2/M期阻滞,诱导HEL细胞向巨核细胞方向分化,且呈剂量效应。Westernblotting检测结果表明L6主要通过激活Caspase-3执行细胞凋亡。小鼠肝组织苏木精-伊红(HE)染色显示肝组织内HEL细胞浸润有所减轻,但L6组减轻的效果更明显,表明其可延缓白血病细胞的转移,且效果优于米哚妥林。结论 双吲哚马来酰亚胺衍生物L6有较强的抗白血病活性,为新型白血病药物的研发提供参考。
[Key word]
[Abstract]
Objective To investigate the effect of bisindolylmaleimide derivative L6 on inducing apoptosis of leukemia cells and its molecular mechanism. Methods MTT assay was used to determinate the killing effect of L6 on HELL, K562, and KG1a cells. Flow cytometry was used to detect the effects of L6 on apoptosis, cell cycle, and differentiation of HEL cells. Western blotting was used to detect the expression of apoptosis-related protein. Finally, the effect of L6 on leukemia mouse was studied in vivo. Results MTT assay showed that L6 had a stronger inhibitory activity against HEL, K562, and KG1a cell lines than the positive control PKC412 compound, with IC50 of (0.05 ±0.03), (0.32 ±0.01), and (0.19 ±0.10) μmol/L, respectively. L6 could induce the apoptosis, G2/M arrest, megakaryocyte differentiation of HEL cells with a dose effect. Western blotting revealed that L6 mainly performed apoptosis by activating Caspase-3, which is an apoptotic executive protein. Hematoxylin-eosin (HE) staining of liver tissue of mice showed a reduction in HEL cell infiltration, but the more significant reduction in group L6 was observed, indicating that L6 could delay the metastasis of leukemia, and its effect was better than that of PKC412. Conclusion Bisindolylmaleimide derivative L6 has a strong anti-leukemia activity, providing new hope for the development of new leukemia drugs.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金资助项目(81872772);国家自然科学基金资助项目(81700169);贵州省科技计划项目(黔科合支撑[2019]2762);贵州省百人计划项目