一种天然植物抗菌液PAMs对人白血病K562细胞的促凋亡作用

初步研究一种天然植物抗菌液(PAMs)对白血病K562细胞的杀伤作用及抗肿瘤分子作用机制。MTT法确定PAMs对K562细胞的增殖抑制作用和具有的浓度和时间依赖性；AO-EB双染、Annexin-FITC/PI流式细胞仪染色观察显示PAMs对K562细胞的杀伤作用与细胞凋亡相关，这也被进一步的分子水平与酶活力检测所证实。FQ-PCR基因表达检测发现，PAMs处理后，K562细胞株中caspase-3、caspase-9、bax等促凋亡相关基因有所上升，而抑制凋亡基因bcl-2表达下降，Western blot法检测到PAMs上调caspase-3的表达量，同时下调抑制凋亡蛋白survivin的表达。这也与caspase酶活性测定结果相一致，PAMs处理后的K562，其caspase-3、-9活性均有所上升。所有这些结果一方面揭示PAMs对白血病K562细胞株具有强的杀伤作用，同时在形态和分子水平揭示其抗肿瘤抑制作用途径与诱导细胞凋亡的密切相关性，为PAMs作为抗白血病抗肿瘤中药的研发提供参考。

Promotion of apoptosis in leukemia K562 cells by natural plant-derived antimicrobial solution(PAMs)

In this study, the leukemia K562 cell line was used as a model to elucidate the anticancer effects and preliminary mechanisms of PAMs. MTT assay showed that PAMs could cause cytotoxicities in K562 cells in dose- and time-dependent manners. AO-EB, Annexin-FITC/PI staining showed that the killing effects of PAMs in K562 cells were related to apoptosis, which was further confirmed by the following molecular and enzymatic assay. The mRNA levels of pro-apoptotic genes caspase-3, caspase-9 and bax were remarkably increased while the anti-apoptotic gene bcl-2 was significantly decreased determined by fluorescent quantitative PCR. Western blotting disclosed that PAMs could up-regulate caspase-3 and down-regulate anti-apoptotic survivin protein expression. The latter was also consistent with the results that PAMs could increase the enzymatic activities of both caspase-3 and caspase-9. All these results suggested that PAMs could effectively inhibit the proliferation of K562 cells and the mechanisms may be closely related to apoptosis induction. The work provides evidence basis for PAMs to be potentially developed as anti-cancer leukemia Chinese medicine.