新合成的7-氮杂异靛蓝抑制A549细胞增殖的作用及可能机制

The Inhibitory Effect and Possible Mechanism of a Novel 7-azaisoindigo on Proliferation of A549 Cells

研究新合成的N^1-(正-丁基)-7-氮杂异靛蓝(N^1-(n-butyl)-7-azaisoindigo,7-AI-b)抑制人非小型肺癌细胞A549的增殖,初步探索其抗肿瘤的机制。以不同浓度的7-AI-b作用于A549,MTT检测与相差显微镜观察相结合,分析7-AI-b对细胞增殖的影响;MDC染色和Western印迹检测自噬标志蛋白(LC3)的表达,研究药物处理后细胞是否发生了自噬。PI单染,流式检测细胞周期的变化;Fluo-3-AM荧光探针检测细胞中Ca^(2+)的含量。结果发现,7-AI-b以时间和剂量依赖性方式抑制细胞的增殖;MDC染色后,自噬小泡增多,LC3的总表达量增加,且由LC3-Ⅰ向LC3-Ⅱ的转变量升高。流式细胞检测发现细胞被阻滞在G_0/G_1期;同时,药物处理后,细胞内Ca^(2+)出现了超载。由此,新合成的7-AI-b有很好的抑制肿瘤细胞增殖的作用,其机制与引起细胞周期阻滞、诱导细胞自噬有关,胞内Ca^(2+)超载也参与了该作用。

The anticancer effect of a newly synthesized 7-azaisoindigo derivative （namely N^1-（n-butyl）-7- azaisoindigo, 7-AI-b） was investigated in human lung cell line A549, and the underlying mechanism of action was analysed. Cell proliferation was assessed by MTT assay; Autophagy induced by 7-AI-b was evaluated by MDC staining and the expression of LC3 by Western blot; Cell cycle was detected by flow cytometry （FCM） following PI staining; The alteration of intracellular Ca^2＋ level was detected following Fluo-3-AM loading. It was found that 7-AI- b inhibited A549 cell proliferation in a dose- and time-dependent way. 7-AI-b-induced autophagy occurred showing enhanced MDC staining, and increased in both expressions of LC3 and conversion of LC3-Ⅰ to LC3-Ⅱ; Cell cycle was arrested in the G0/G1 phase; Meanwhile, 7-AI-b was found to increase intracellular Ca^2＋ level. Generally speaking, the novel compound 7-AI-b could inhibit the proliferation of cancer cells effectively. Its mechanism might be related to arrest cell cycle and induce cell autophagy. The intracellular Ca^2＋ overloading was involved in cell death process.