卤地菊乙醇提取物W40单体诱导GLC-82细胞凋亡的分子机制研究

doi:10.13523/j.cb.20170801

中国生物工程杂志

2017, Vol. 37

Issue (8): 1-7 DOI: 10.13523/j.cb.20170801

研究报告

卤地菊乙醇提取物W40单体诱导GLC-82细胞凋亡的分子机制研究

代立婷¹, 吴忠南², 黄翔¹, 杨杰¹, 曾慧兰¹, 王国才², 蒋建伟¹

1. 暨南大学医学院广州 510632;
2. 暨南大学药学院中药及天然产物研究所广州 510632

Molecular Mechanism of Inducing GLC-82 Cells Apoptosis by Ethanol Extract from Wedelia prostrate(Hook.et Arn.) Hemsl

DAI Li-ting¹, WU Zhong-nan², HUANG Xiang¹, YANG Jie¹, ZENG Hui-lan¹, WANG Guo-cai², JIANG Jian-wei¹

1. Department of Biochemistry, Medical College, Jinan University, Guangzhou 510632, China;
2. Institute of Traditional Chinese Medicine and Nature Products, Jinan University, Guangzhou 510630, China

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摘要： 目的：探讨卤地菊Wedelia prostrate（Hook.et Arn.） Hemsl.的乙醇提取物W40单体对非小细胞肺癌GLC-82细胞的抗肿瘤作用分子机制。方法：通过MTT和克隆形成抑制实验检测W40对非小细胞肺癌GLC-82细胞增殖的影响；通过细胞划痕实验检测W40对细胞迁移的影响；通过Annexin V-FITC/PI双染法检测W40对细胞凋亡的诱导；通过Western blotting分析细胞增殖和凋亡相关蛋白的水平。结果：MTT实验表明，W40对GLC-82细胞有较为明显的细胞毒作用；克隆形成抑制实验表明，W40可以显著抑制GLC-82细胞增殖，且抑制程度呈现浓度依赖效应；细胞划痕实验表明，W40能够在一定程度上抑制GLC-82细胞的迁移能力；Annexin V-FITC/PI双染的结果显示，随着药物浓度的增加，细胞凋亡率逐渐增加；Western blotting结果表明，随着药物浓度的增加，p-Stat3蛋白的表达水平下降，Stat3下游蛋白Bcl-2、Mcl-1的表达减少。同时，促凋亡蛋白Bax的表达增多，并伴随Caspase-9、Caspase-3活化及多聚ADP-核糖聚合酶PARP酶切失活；p-BRAF蛋白的表达水平下降，p-BRAF下游蛋白p-MEK、p-ERK表达减少。结论：W40通过抑制BRAF/MAPK/ERK及Stat3信号通路来诱导细胞凋亡。

关键词： 肺癌; Stat3; 卤地菊; 细胞凋亡; BRAF/MAPK/ERK信号通路

Abstract: Objective:To study the antitumor mechanism of W40, a monomer purified from Wedelia prostrate (Hook.et Arn.) Hemsl. Methods:The effects of W40 on the cell proliferative of GLC-82 cells were detected by MTT assay and colony formation assay. The migratory abilities of GLC-82 cells were observed by wound healing assay. Cell apoptosis was evaluated by Annexin V-FITC/PI staining analysis.The levels of apoptosis-relative proteins and cell proliferation-related proteins, such as Caspase-3, PARP, Stat3 and ERK, were detected by Western blotting. Results:MTT assay showed that W40 had a significant cytotoxic effect on non-small cell lung cancer GLC-82 cells. Colony formation assays showed that W40 significantly inhibited GLC-82 cells proliferation.The migration of GLC-82 cells was inhibited by W40 in a dose-dependent manner. Flow cytometry showed that the apoptotic rate increased gradually in a concentration-dependent manner. W40 down-regulated Stat3 as decreasing p-Stat3 and downstream proteins of Bcl-2 and Mcl-1. At the same time, W40 up-regulated the expression of pro-apoptotic protein Bax, and increased the cleavaged Caspase-9,Caspase-3 and PARP. W40 also down-regulated BRAF/MAPK/ERK signal pathway as decreasing p-BRAF, p-MEK and p-ERK. Conclusions:W40 induced apoptosis by inhibiting BRAF/MAPK/ERK and Stat3 signaling pathways.

Key words: Wedelia prostrate Apoptosis Stat3 Lung cancer BRAF/MAPK/ERK signaling pathways

收稿日期: 2017-05-23 出版日期: 2017-08-25

ZTFLH:

Q819

基金资助: 广东省自然科学基金（2014A030313356）、广东省医学科研基金（2015130164321691）资助项目

通讯作者: 蒋建伟 E-mail: jjw703@163.com

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引用本文:

代立婷, 吴忠南, 黄翔, 杨杰, 曾慧兰, 王国才, 蒋建伟. 卤地菊乙醇提取物W40单体诱导GLC-82细胞凋亡的分子机制研究[J]. 中国生物工程杂志, 2017, 37(8): 1-7.

DAI Li-ting, WU Zhong-nan, HUANG Xiang, YANG Jie, ZENG Hui-lan, WANG Guo-cai, JIANG Jian-wei. Molecular Mechanism of Inducing GLC-82 Cells Apoptosis by Ethanol Extract from Wedelia prostrate(Hook.et Arn.) Hemsl. China Biotechnology, 2017, 37(8): 1-7.

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https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170801 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I8/1

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