Samarghandian Saeed, Nezhad Mohsen Azimi, Mohammadi Gholamreza
Department of Basic Medical sciences, Neyshabur University of Medical Sciences, Neyshabur, Iran.
Anticancer Agents Med Chem. 2014;14(6):901-9. doi: 10.2174/1871520614666140209144042.
Honey is reported to contain various compounds such as antioxidants. Chrysin is a natural and biologically active compound extracted from honey. It possesses antioxidant properties and promotes cell death by perturbing cell cycle progression. We focused on the possible role that chrysin may act as a potential anticancer agent, and tested its biological activity and possible mechanisms in the human lung adenocarcinoma epithelial cell line.
MATERIALS & METHODS: Antiproliferative effect of honey and chrysin were determined by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay; DNA fragmentation was determined by gel electrophoresis assay; apoptosis was detected by flow cytometer; apoptosis-related gene expression was detected by reverse transcription polymerase chain reaction assay; and activation of caspase-3 and caspase-9 were evaluated by a colorimetric assay; Bax and Bcl-2 protein expression were also analysed by western blotting.
The results revealed that the cell viability decreased in a concentration- and time- dependent manner in the malignant cells treated with honey and chrysin in comparison with the nonmalignant cells. The IC50 values of honey against A549 cells were determined 15 ± 0.05% and 8 ± 0.05 % after 48 and 72h, respectively. The IC50 dose of chrysin was determined to be 49.2 ± 0.6 and 38.7 ± 0.8 μM at 48 and 72 h, respectively. Reactivity with Annexin V fluorescence antibody and propidium iodide showed that chrysin induced apoptosis in the lung cancer cells (p<0.001). Moreover, chrysin treatment resulted in the activation of caspase-3 and - 9 and an increase in the Bax/Bcl-2 ratio (p<0.01). Bax protein expression was increased but Bcl-2 protein expression decreased in chrysin-treated cells .Chrysin inhibits the growth of the lung cancer cells by inducing cancer cell apoptosis via the regulation of the Bcl-2 family and also activation of caspase-3 and -9, which may, in part, explain its anticancer activity.
This study shows that chrysin could also be considered as a promising chemotherapeutic agent and anticancer activity in treatment of the lung cancer cells in future.
据报道，蜂蜜含有多种化合物，如抗氧化剂。白杨素是从蜂蜜中提取的一种天然生物活性化合物。它具有抗氧化特性，并通过干扰细胞周期进程促进细胞死亡。我们重点研究了白杨素作为潜在抗癌剂的可能作用，并在人肺腺癌上皮细胞系中测试了其生物活性及可能机制。
采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定蜂蜜和白杨素的抗增殖作用；通过凝胶电泳法测定DNA片段化；用流式细胞仪检测细胞凋亡；采用逆转录聚合酶链反应法检测凋亡相关基因表达；通过比色法评估半胱天冬酶-3和半胱天冬酶-9的激活情况；还通过蛋白质印迹法分析Bax和Bcl-2蛋白表达。
结果显示，与非恶性细胞相比，用蜂蜜和白杨素处理的恶性细胞的细胞活力呈浓度和时间依赖性下降。蜂蜜作用于A549细胞48小时和72小时后的IC50值分别为15±0.05%和8±0.05%。白杨素在48小时和72小时时的IC50剂量分别为49.2±0.6和38.7±0.8μM。与膜联蛋白V荧光抗体和碘化丙啶反应表明，白杨素可诱导肺癌细胞凋亡（p<0.001）。此外，白杨素处理导致半胱天冬酶-3和-9激活，Bax/Bcl-2比值升高（p<0.01）。在白杨素处理的细胞中，Bax蛋白表达增加而Bcl-2蛋白表达减少。白杨素通过调节Bcl-2家族以及激活半胱天冬酶-3和-9诱导癌细胞凋亡，从而抑制肺癌细胞生长，这可能部分解释了其抗癌活性。
本研究表明，白杨素在未来治疗肺癌细胞方面也可被视为一种有前景的化疗药物并具有抗癌活性。
