垂花水冬瓜二氯甲烷根提取物诱导 MDA-MB-231 三阴性乳腺癌细胞凋亡。

Dillenia suffruticosa dichloromethane root extract induced apoptosis towards MDA-MB-231 triple-negative breast cancer cells.

机构信息

Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

出版信息

J Ethnopharmacol. 2016 Jul 1;187:195-204. doi: 10.1016/j.jep.2016.04.048. Epub 2016 Apr 27.

DOI:10.1016/j.jep.2016.04.048

PMID:27131434

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Dillenia suffruticosa is traditionally used for treatment of cancerous growth including breast cancer in Malaysia.

AIM OF THE STUDY

Dillenia suffruticosa is a well-known medicinal plant in Malaysia for the treatment of cancer. Nevertheless, no study has been reported the cytotoxicity of this plant towards MDA-MB-231 triple-negative breast cancer cells. The present study was designed to investigate the mode of cell death and signalling pathways of MDA-MB-231 cells treated with dichloromethane Dillenia suffruticosa root extract (DCM-DS).

METHODS

Extraction of Dillenia suffruticosa root was performed by the use of sequential solvent procedure. The cytotoxicity of DCM-DS was determined by using MTT assay. The mode of cell death was evaluated by using an inverted light microscope and flow cytometry analysis using Annexin-V/PI. Cell cycle analysis and measurement of reactive oxygen species level were performed by using flow cytometry. The cells were treated with DCM-DS and antioxidants α-tocopherol or ascorbic acid to evaluate the involvement of ROS in the cytotoxicity of DCM-DS. Effect of DCM-DS on the expression of antioxidant, apoptotic, growth, survival genes and proteins were analysed by using GeXP-based multiplex system and Western blot, respectively. The cytotoxicity of compounds isolated from DCM-DS was evaluated towards MDA-MB-231 cells using MTT assay.

RESULTS

DCM-DS induced apoptosis, G2/M phase cell cycle arrest and oxidative stress in MDA-MB-231 cells. The induction of apoptosis in MDA-MB-231 cells by DCM-DS is possibly due to the activation of pro-apoptotic JNK1 and down-regulation of anti-apoptotic ERK1, which in turn down-regulates anti-apoptotic BCL-2 to increase the BAX/BCL-2 ratio to initiate the mitochondrial apoptotic pathway. The cell cycle arrest in DCM-DS-treated MDA-MB-231 cells is possibly via p53-independent but p21-dependent pathway. A total of 3 triterpene compounds were isolated from DCM-DS. Betulinic acid appears to be the most major and most cytotoxic compound in DCM-DS.

CONCLUSION

The data suggest the potential application of DCM-DS in the treatment of triple-negative breast cancer.

摘要

民族药理学相关性

在马来西亚，杜茎山被传统用于治疗包括乳腺癌在内的癌性生长。

研究目的

杜茎山是马来西亚一种著名的药用植物，用于治疗癌症。然而，目前尚无研究报道该植物对 MDA-MB-231 三阴性乳腺癌细胞的细胞毒性。本研究旨在探讨二氯甲烷杜茎山根提取物（DCM-DS）处理 MDA-MB-231 细胞的细胞死亡方式和信号通路。

方法

采用连续溶剂法提取杜茎山根。用 MTT 法测定 DCM-DS 的细胞毒性。用倒置显微镜和 Annexin-V/PI 流式细胞术分析评估细胞死亡方式。用流式细胞术进行细胞周期分析和活性氧（ROS）水平测定。用 DCM-DS 和抗氧化剂α-生育酚或抗坏血酸处理细胞，以评估 ROS 在 DCM-DS 细胞毒性中的作用。用 GeXP 多重系统和 Western blot 分别分析 DCM-DS 对抗氧化、凋亡、生长、存活基因和蛋白表达的影响。用 MTT 法评价从 DCM-DS 中分离得到的化合物对 MDA-MB-231 细胞的细胞毒性。

结果

DCM-DS 诱导 MDA-MB-231 细胞凋亡、G2/M 期细胞周期阻滞和氧化应激。DCM-DS 诱导 MDA-MB-231 细胞凋亡可能是通过激活促凋亡 JNK1 和下调抗凋亡 ERK1，从而下调抗凋亡 BCL-2 增加 BAX/BCL-2 比值启动线粒体凋亡途径。DCM-DS 处理的 MDA-MB-231 细胞的细胞周期阻滞可能是通过 p53 非依赖性但 p21 依赖性途径。从 DCM-DS 中分离得到 3 种三萜类化合物。白桦脂酸似乎是 DCM-DS 中最主要和最具细胞毒性的化合物。