SMT-A07, a 3-(Indol-2-yl) indazole derivative, induces apoptosis of leukemia cells in vitro
SMT-A07, a 3-(Indol-2-yl) indazole derivative, induces apoptosis of leukemia cells in vitro
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SMT-A07，一种 3-(Indol-2-yl) 吲唑衍生物，体外诱导白血病细胞凋亡
DOI：
10.1007/s11010-010-0554-y
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发表时间：
2010-12-01
影响因子：
4.3
通讯作者：
Yang, Bo
中科院分区：
文献类型：
--
作者：
Qian, Shijing;Cao, Ji;Yang, Bo
N-（2-（1H-吲唑-3-基）-1H-吡咯并[3，2-B]吡啶-5-基）-4-氯-N-甲基苯甲酰胺（SMT-A07）是一种新的3-（吲哚-2-基）吲唑衍生物。本实验研究了SMT-A07的体外抗肿瘤活性及对人白血病HL 60和NB 4细胞的诱导凋亡作用。MTT法结果表明，SMT-A07是一种潜在的高效抗肿瘤化合物，对5种白血病细胞的IC 50值为0.09 ~ 1.19 μM。流式细胞仪检测发现，SMT-A07作用HL 60细胞24 h后，细胞凋亡率由6.88%增加到49.72%，NB 4细胞24 h后，细胞凋亡率由8.72%增加到56.28%。琼脂糖凝胶电泳显示细胞暴露于SMT-A07后出现DNA片段化。SMT-A07孵育后，DAPI染色显示存在DNA片段化和核周凋亡小体。JC-1染色结果显示，SMT-A07对HL 60和NB 4细胞的Δ λ m均无明显影响。并通过Western blotting检测其细胞毒作用相关蛋白，探讨其细胞毒作用机制。SMT-A07暴露引起半胱天冬酶原-8、半胱天冬酶原-3、Bid、PARP的下调和裂解以及裂解的半胱天冬酶-8、裂解的半胱天冬酶-3、PARP（裂解片段）的上调。此外，泛半胱天冬酶抑制剂BOC-D-FMK的存在阻止了细胞的半胱天冬酶-3活化、PARP裂解和随后的凋亡。我们的研究表明SMT-A07具有明显的抗肿瘤活性和广泛的抗白血病谱，并且SMT-A07可以诱导HL 60和NB 4细胞凋亡，激活caspase级联反应，值得进一步研究。
N-(2-(1H-indazol-3-yl)-1H-pyrrolo[3,2-b]pyridin-5-yl)-4-chloro-N-methylbenzamide (SMT-A07) is a novel 3-(Indol-2-yl) indazole derivative. The anticancer activities in vitro and the cell apoptosis-induction abilities of SMT-A07 on human leukemia HL60 and NB4 cell lines were investigated in this study. The results of MTT assay showed SMT-A07 was a potential and highly efficient antitumor compound with IC50values ranging from 0.09 to 1.19 μM in five leukemia cell lines. SMT-A07 treatment for 24 h caused the increment of apoptosis rate from 6.88 to 49.72% in HL60 cells and from 8.72 to 56.28% in NB4 cells by flow cytometry analysis. Agarose gel electrophoresis showed DNA fragmentation that appeared after cells were exposed to SMT-A07. After SMT-A07 incubation, DAPI staining revealed the presence of DNA fragmentation, and perinuclear apoptotic body. SMT-A07 also resulted in a loss of ΔΨm in both HL60 and NB4 cells by JC-1 staining. Moreover, apoptosis-related proteins were examined by western blotting to explore the mechanism of its cytotoxicity. SMT-A07 exposure caused down-regulation and cleavage of procaspase-8, procaspase-3, Bid, PARP and up-regulation of cleaved caspase-8, cleaved caspase-3, PARP (Cleaved Fragment). In addition, the presence of pan-caspase inhibitor BOC-D-FMK prevented cells from caspase-3 activation, PARP cleavage, and subsequent apoptosis. Our study demonstrates that SMT-A07 displays an apparent antitumor activity with extensive anti-leukemia spectrum, and SMT-A07 can induce the apoptosis of HL60 and NB4 cells activation of the caspase cascade, which deserves further development.