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[目的]观察苦瓜蛋白对A549非小细胞肺癌细胞凋亡的影响,探讨苦瓜蛋白对PI3K/AKT细胞信号传导通路中相关蛋白分子的作用影响。[方法]以梯度浓度苦瓜蛋白处理A549细胞48h,流式细胞术(TUNEL方法)检测A549细胞凋亡;10μg/ml苦瓜蛋白处理A549细胞,分别作用不同时间(0h、6h、12h、24h、48h、72h)后,提取细胞总蛋白,Westernblot检测AKT信号通路中AKT及p-AKT等信号分子表达的变化。[结果]苦瓜蛋白可以明显诱导A549细胞凋亡,以10μg/ml苦瓜蛋白处理组A549细胞凋亡率最高(50.15%)。苦瓜蛋白处理A549细胞12h后,p-AKT及AKT表达明显下调,呈现时间—效应依赖关系。[结论]苦瓜蛋白可以诱导A549细胞发生显著凋亡,其机制可能是部分通过抑制AKT蛋白的表达及其磷酸化,抑制AKT信号传导通路,导致肿瘤细胞凋亡。
[Objective] To observe the effect of bitter melon on the apoptosis of A549 non-small cell lung cancer cells and explore the effect of bitter melon on the related protein molecules in PI3K / AKT cell signal transduction pathway. [Methods] The A549 cells were treated with gradient concentration of bitter melon for 48 hours and the apoptosis of A549 cells was detected by flow cytometry (TUNEL method). A549 cells were treated with 10μg / ml bitter melon for different time (0h, 6h, 12h, 24h, 48h , 72h), total cellular protein was extracted and the expression of AKT and p-AKT in AKT signaling pathway was detected by Western blot. [Result] Momordin could obviously induce the apoptosis of A549 cells. The apoptosis rate of A549 cells treated with 10μg / ml Momordicin was the highest (50.15%). The expression of p-AKT and AKT was significantly down-regulated after treated with Momordin for 12 hours in A549 cells, showing a time-dependent relationship. [Conclusion] Momordica charantia protein can induce significant apoptosis in A549 cells, which may be partly through inhibiting AKT protein expression and phosphorylation, inhibiting AKT signaling pathway and leading to tumor cell apoptosis.