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利用流式细胞术技术研究离子液体-西番莲果提取物对人乳腺癌(MCF-7)细胞系的抗增殖活性。

Antiproliferative activity of ionic liquid-graviola fruit extract against human breast cancer (MCF-7) cell lines using flow cytometry techniques.

机构信息

Biotechnology Engineering, Kulliyyah of Engineering, International Islamic University, Malaysia (IIUM), P. O. Box 10, Gombak, 50728, Kuala Lumpur, Malaysia.

International Institute for Halal Research and Training (INHART), Level 3, KICT Building, International Islamic University Malaysia (IIUM), Jalan Gombak, 53100, Kuala Lumpur, Malaysia.

出版信息

J Ethnopharmacol. 2019 May 23;236:466-473. doi: 10.1016/j.jep.2019.03.003. Epub 2019 Mar 7.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Medicinal plants have been used for ages by indigenous communities around the world to help humankind sustain its health. Graviola (Annona muricata), also called soursop, is a member of the Annonaceae family and is an evergreen plant that is generally distributed in tropical and subtropical areas of the world. Graviola tree has a long history of traditional use due to its therapeutic potential including anti-inflammatory, antimicrobial, antioxidant, insecticide and cytotoxic to tumor cells.

AIM OF THE STUDY

This study aimed to investigate the in vitro antiproliferative effects and apoptotic events of the ionic liquid extract of Graviola fruit (IL-GFE) on MCF-7 breast cancer cells and their cytokinetics behaviour to observe their potential as a therapeutic alternative in cancer treatment.

MATERIALS AND METHODS

The cell viability assay of the extract was measured using tetrazolium bromide (MTT assay) to observe the effects of Graviola fruit extract. Then the cytokinetics behaviour of MCF-7 cells treated with IL-GFE is observed by plotting the growth curve of the cells. Additionally, the cell cycle distribution and apoptosis mechanism of IL-GFE action on MCF-7 cancer cells were observed by flow cytometry.

RESULTS

IL-GFE exhibited anti-proliferative activity on MCF-7 with the IC50 value of 4.75 μg/mL, compared to Taxol with an IC50 value of 0.99 μg/mL. IL- GFE also reduced the number of cell generations from 3.71 to 1.67 generations compared to 2.18 generations when treated with Taxol. Furthermore, the anti-proliferative activities were verified when the growth rate was decreased dynamically from 0.0077 h to 1 to 0.0035 h-1. Observation of the IL-GFE-treated MCF-7 under microscope demonstrated detachment of cells and loss of density. The growth inhibition of the cells by extracts was associated with cell cycle arrest at the G0/G1 phase, and phosphatidylserine externalisation confirms the anti-proliferation through apoptosis.

CONCLUSIONS

ionic liquid Graviola fruit extract affect the cytokinetics behaviour of MCF-7 cells by reducing cell viability, induce apoptosis and cell cycle arrest at the G0/G1 phase.

摘要

民族药理学相关性

药用植物被世界各地的土著社区使用了很长时间,以帮助人类维持健康。刺果番荔枝(Annona muricata),也称为刺果番荔枝,是番荔枝科的一员,是一种常绿植物,通常分布在世界的热带和亚热带地区。由于其治疗潜力,包括抗炎、抗菌、抗氧化、杀虫和对肿瘤细胞的细胞毒性,刺果番荔枝树有着悠久的传统用途历史。

研究目的

本研究旨在研究离子液体提取的刺果番荔枝(IL-GFE)对 MCF-7 乳腺癌细胞的体外增殖抑制作用和细胞凋亡事件及其细胞动力学行为,以观察其作为癌症治疗替代疗法的潜力。

材料和方法

使用四唑溴盐(MTT 测定)测量提取物的细胞活力,以观察刺果番荔枝提取物的影响。然后通过绘制细胞生长曲线观察用 IL-GFE 处理的 MCF-7 细胞的细胞动力学行为。此外,通过流式细胞术观察 IL-GFE 对 MCF-7 癌细胞的细胞周期分布和凋亡机制。

结果

与 IC50 值为 0.99μg/mL 的紫杉醇相比,IL-GFE 对 MCF-7 表现出抗增殖活性,IC50 值为 4.75μg/mL。与紫杉醇处理时的 2.18 代相比,IL-GFE 还将细胞世代数从 3.71 减少到 1.67 代。此外,当生长速率从 0.0077 h 动态降低到 1 到 0.0035 h-1 时,抗增殖活性得到验证。在显微镜下观察到用 IL-GFE 处理的 MCF-7,细胞分离和密度丧失。提取物对细胞的生长抑制与细胞周期停滞在 G0/G1 期有关,并且磷脂酰丝氨酸的外化证实了通过细胞凋亡的增殖抑制。

结论

离子液体刺果番荔枝果提取物通过降低细胞活力、诱导细胞凋亡和细胞周期停滞在 G0/G1 期来影响 MCF-7 细胞的细胞动力学行为。

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