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巴豆叶提取物对肺癌细胞系A549的细胞毒性和促凋亡活性。

Cytotoxic and pro-apoptotic activities of leaf extract of Croton bonplandianus Baill. against lung cancer cell line A549.

作者信息

Bhavana J, Kalaivani M K, Sumathy A

出版信息

Indian J Exp Biol. 2016 Jun;54(6):379-85.

Abstract

The acetone extract (AcE) of the Croton bonplandianus Baill., an exotic weed of the Euphorbiaceae family was studied for cytotoxicity, apoptosis, cell cycle arrest in A549 cell line and antioxidant capacities using MTT assay, acridine orange/ethidium bromide (AO/EB staining), cell cycle analysis and DPPH radical scavenging assay respectively. Based on the cytotoxic activity, the extract was tested for the apoptotic effect using AO/EB and Hoechst 33258 staining. The apoptosis was characterized by chromatin condensation and DNA fragmentation. Further, to determine the stage of cell death, cell cycle analysis was performed by flow cytometry and AcE was found to arrest G2/M phase in a dose dependent manner. The number of cells in G2/M phase increases with concurrent accumulation of cells in sub G₀/G₁phase indicates the induction of apoptosis at G2M phase. The free radical scavenging activity of the AcE against DPPH was considerably significant. The cytotoxic, apoptotic and antioxidant effect of the AcE could be well correlated with the presence of potent free radical scavenging secondary metabolites such as phenols (43 ± 0.05 µg/mL), flavonoids (3.5 ± 0.07 µg/mL) and tannin (0.36 ± 0.1 µg/mL). Our study has shown that A549 cells were more sensitive to AcE with an IC₅₀ of 15.68 ± 0.006 µg/mL compared to the standard drug 2.20 ± 0.008 µg/mL (cisplatin). The results suggest that Croton bonplandianus could serve as a potential source of alternative therapeutic agent for treating cancer. Further research is required to isolate the active principle compound and determination of its anticancer property.

摘要

对大戟科外来杂草巴豆(Croton bonplandianus Baill.)的丙酮提取物(AcE)进行了研究,分别采用MTT法、吖啶橙/溴化乙锭(AO/EB染色)、细胞周期分析和DPPH自由基清除试验,检测其对A549细胞系的细胞毒性、凋亡、细胞周期阻滞及抗氧化能力。基于细胞毒性活性,使用AO/EB和Hoechst 33258染色检测提取物的凋亡效应。凋亡的特征为染色质浓缩和DNA片段化。此外,为确定细胞死亡阶段,通过流式细胞术进行细胞周期分析,发现AcE以剂量依赖性方式阻滞G2/M期。G2/M期细胞数量增加,同时亚G₀/G₁期细胞积累,表明在G2M期诱导了凋亡。AcE对DPPH的自由基清除活性相当显著。AcE的细胞毒性、凋亡和抗氧化作用与酚类(43±0.05μg/mL)、黄酮类(3.5±0.07μg/mL)和单宁(0.36±0.1μg/mL)等具有强大自由基清除能力的次生代谢产物的存在密切相关。我们的研究表明,与标准药物顺铂(2.20±0.008μg/mL)相比,A549细胞对AcE更敏感,IC₅₀为15.68±0.006μg/mL。结果表明,巴豆可能是治疗癌症的潜在替代治疗剂来源。需要进一步研究分离活性成分并确定其抗癌特性。

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