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白花老鸦嘴水提取物在人乳腺癌细胞中产生自由基中间体并诱导细胞周期停滞。

Radical intermediate generation and cell cycle arrest by an aqueous extract of Thunbergia Laurifolia Linn. In human breast cancer cells.

作者信息

Jetawattana Suwimol, Boonsirichai Kanokporn, Charoen Savapong, Martin Sean M

机构信息

Research and Development Group, Thailand Institute of Nuclear Technology (Public Organization), Nakhon Nayok, Thailand E-mail :

出版信息

Asian Pac J Cancer Prev. 2015;16(10):4357-61. doi: 10.7314/apjcp.2015.16.10.4357.

Abstract

Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an IC50 value of 843 μg/ml. Treatments with extract for 24 h at 250 μg/ml or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

摘要

黑叶老鸦嘴是泰国传统医学中最常见的植物之一,可用于治疗多种病症,包括癌症。然而,黑叶老鸦嘴及其成分的抗肿瘤活性在分子水平上从未有过报道,无法支持民间说法。本研究旨在调查黑叶老鸦嘴水提取物对人乳腺癌细胞的抗肿瘤作用及其可能的作用机制。从黑叶老鸦嘴的干燥叶片中制备了水粗提取物。采用福林-西奥尔特试剂比色法测定总酚含量。通过MTT还原试验、细胞生长抑制试验、克隆形成细胞存活试验和流式细胞术分析,评估了其对人乳腺腺癌MCF-7细胞的抗增殖和细胞周期作用。使用电子顺磁共振光谱检测提取物产生的自由基。人乳腺腺癌MCF-7细胞暴露于黑叶老鸦嘴水提取物后,细胞生长、克隆形成细胞存活和细胞活力呈浓度依赖性降低,IC50值为843μg/ml。在250μg/ml或更高浓度下用提取物处理24小时可诱导细胞周期停滞,表现为细胞周期G1期细胞群体显著增加,S期显著减少。在高pH和近中性pH条件下均观察到水提取物产生自由基中间体的能力。研究结果表明,黑叶老鸦嘴对所选乳腺癌细胞的抗肿瘤生物活性可能是由于诱导G1期细胞周期停滞。提取物高浓度时相关的细胞毒性和细胞周期扰动,部分可由提取物中的总酚含量以及产生自由基中间体以调节细胞增殖信号的能力来解释。

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