芳樟醇氧化物 2 通过 ROS 介导的线粒体途径诱导皮肤表皮癌细胞凋亡。

Aromadendrene oxide 2, induces apoptosis in skin epidermoid cancer cells through ROS mediated mitochondrial pathway.

机构信息

School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore 632 014, India.

Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600 036, India.

出版信息

Life Sci. 2018 Mar 15;197:19-29. doi: 10.1016/j.lfs.2018.01.029. Epub 2018 Jan 31.

DOI:10.1016/j.lfs.2018.01.029

PMID:29407546

Abstract

AIMS

Aromadendrene oxide 2 (AO-(2)) is an oxygenated sesquiterpene naturally found as a chemical component of essential oils. In the present study anticancer activity of AO-(2) has been investigated on A431 human epidermoid cancer and precancerous HaCaT cells.

MATERIAL AND METHODS

Cell viability was used to detect cytotoxic activity. Mechanism of cell death induced by AO-(2) treatments was studied using Annexin V-FITC/PI binding, cell cycle analysis, measurement of MMP and ROS generation by flow cytometry. Expression of apoptosis related proteins was investigated by western blot.

KEY FINDINGS

AO-(2) inhibited the growth and colony formation ability of A431 and HaCaT cells in concentration dependent manner. It induced cell cycle arrest at G0/G1 phase and apoptosis through intracellular ROS accumulation. Inhibition of intracellular ROS by ascorbic acid and N-acetyl cysteine treatment completely blocked apoptotic effect. N-acetyl cysteine treatment significantly reversed G0/G1 arrest induced by AO-(2). AO-(2) treatment caused loss of mitochondrial membrane potential (ΔΨm), increase in Bax/Bcl-2 ratios, cytochrome c release, activation of caspases (cleaved caspase-3 and caspase-9) and PARP cleavage. AO-(2) also significantly inhibited the growth of multicellular tumor spheroids of A431 and HaCaT cells.

SIGNIFICANCE

The results of the present study reveals that AO-(2) a chemical component of essential oils induces apoptosis in A431 and HaCaT cells.

摘要

目的

芳樟醇氧化物 2（AO-(2)）是一种含氧倍半萜烯，天然存在于精油的化学成分中。在本研究中，研究了 AO-(2) 对 A431 人表皮癌细胞和癌前 HaCaT 细胞的抗癌活性。

材料和方法

使用细胞活力检测来检测细胞毒性活性。通过 Annexin V-FITC/PI 结合、细胞周期分析、流式细胞术测量 MMP 和 ROS 生成来研究 AO-(2) 处理诱导细胞死亡的机制。通过 Western blot 研究凋亡相关蛋白的表达。

主要发现

AO-(2) 以浓度依赖的方式抑制 A431 和 HaCaT 细胞的生长和集落形成能力。它通过细胞内 ROS 积累诱导细胞周期停滞在 G0/G1 期和凋亡。抗坏血酸和 N-乙酰半胱氨酸处理抑制细胞内 ROS 完全阻断了凋亡作用。N-乙酰半胱氨酸处理显著逆转了 AO-(2) 诱导的 G0/G1 期阻滞。AO-(2) 处理导致线粒体膜电位（ΔΨm）丧失、Bax/Bcl-2 比值增加、细胞色素 c 释放、半胱天冬酶（裂解 caspase-3 和 caspase-9）激活和 PARP 裂解。AO-(2) 还显著抑制了 A431 和 HaCaT 细胞的多细胞肿瘤球体的生长。