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海洋微藻角毛藻的粗乙酸乙酯提取物可诱导MDA-MB-231乳腺癌细胞凋亡。

Crude ethyl acetate extract of marine microalga, Chaetoceros calcitrans, induces Apoptosis in MDA-MB-231 breast cancer cells.

作者信息

Goh Su Hua, Alitheen Noorjahan Banu Mohamed, Yusoff Fatimah Md, Yap Swee Keong, Loh Su Peng

机构信息

Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia.

Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM Serdang, Selangor, Malaysia.

出版信息

Pharmacogn Mag. 2014 Jan;10(37):1-8. doi: 10.4103/0973-1296.126650.

Abstract

BACKGROUND

Marine brown diatom Chaetoceros calcitrans and green microalga Nannochloropsis oculata are beneficial materials for various applications in the food, nutraceutical, pharmaceutical and cosmeceutical industries.

OBJECTIVE

This study investigated cytotoxicity of different crude solvent extracts from C. calcitrans and N. oculata against various cancer cell lines.

MATERIALS AND METHODS

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was carried out to screen the cytotoxic effects of hexane (Hex), dichloromethane (DCM), ethyl acetate, and methanol extract from C. calcitrans and N. oculata toward various cancer cell lines. Flow cytometry cell cycle was used to determine the cell cycle arrest while the mode of cell death was investigated through acridine orange/propidium iodide (AOPI) staining, Annexin V-Fluorescein Isothiocyanate (FITC) and Terminal deoxynucleotidyl transferase-mediated d-UTP Nick End Labeling (TUNEL) assays. Expression profile of apoptotic and proliferative-related genes was then determined using the multiplex gene expression profiler (GeXP).

RESULTS

Crude ethyl acetate (CEA) extract of C. calcitrans inhibited growth of MDA-MB-231 cells, with IC50 of 60 μg/mL after 72 h of treatment. Further studies were conducted to determine the mode of cell death at various concentrations of this extract: 30, 60 and 120 μg/mL. The mode of cell death was mainly apoptosis as shown through apoptosis determination test. The expression data from GeXP showed that caspase-4 was upregulated while B-cell leukemia/lymphoma 2(Bcl-2) was down regulated. Thus, caspase-4 induction endoplasmic reticulum death pathway is believed to be one of the mechanisms underlying the induction of apoptosis while Bcl-2 induced S and G2/M cell cycle phase arrest in MDA-MB-231 cells.

CONCLUSION

CEA extract of C. calcitrans showed the highest cytotoxicity on MDA-MB-231 via apoptosis.

摘要

背景

海洋棕色硅藻角毛藻(Chaetoceros calcitrans)和绿色微藻眼点拟微球藻(Nannochloropsis oculata)是食品、营养保健品、制药和药妆品行业各种应用的有益材料。

目的

本研究调查了角毛藻和眼点拟微球藻不同粗溶剂提取物对各种癌细胞系的细胞毒性。

材料与方法

采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法筛选角毛藻和眼点拟微球藻的己烷(Hex)、二氯甲烷(DCM)、乙酸乙酯和甲醇提取物对各种癌细胞系的细胞毒性作用。采用流式细胞术细胞周期分析法确定细胞周期阻滞,同时通过吖啶橙/碘化丙啶(AOPI)染色、膜联蛋白V-异硫氰酸荧光素(FITC)和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)试验研究细胞死亡方式。然后使用多重基因表达谱仪(GeXP)确定凋亡和增殖相关基因的表达谱。

结果

角毛藻的粗乙酸乙酯(CEA)提取物抑制了MDA-MB-231细胞的生长,处理72小时后IC50为60μg/mL。进一步研究确定了该提取物在30、60和120μg/mL不同浓度下的细胞死亡方式。通过凋亡测定试验表明,细胞死亡方式主要为凋亡。GeXP的表达数据显示,半胱天冬酶-4上调,而B细胞白血病/淋巴瘤2(Bcl-2)下调。因此,半胱天冬酶-4诱导内质网死亡途径被认为是诱导凋亡的机制之一,而Bcl-2诱导MDA-MB-231细胞的S期和G2/M期细胞周期阻滞。

结论

角毛藻的CEA提取物通过凋亡对MDA-MB-231表现出最高的细胞毒性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5f4/3969653/b735a65470c4/PM-10-1-g002.jpg

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