齐墩果酸诱导人非小细胞肺癌A549细胞凋亡的作用机制研究  

The Molecular Mechanism of Oleanolic Acid Induced the Apoptosis of Non-small Cells Lung Cancer A549 Cell Line

作　　者：夏荣木 生欣[1] 徐先林[1] 余春波[1] 陆红玲[1] Xia Rongmu;Sheng Xin;Xu Xianlin;Yu Chunbo;Lu Hongling(Zunyi Medical College, Zunyi, 56300)

机构地区：[1]遵义医学院

出　　处：《基因组学与应用生物学》2018年第6期2757-2763,共7页Genomics and Applied Biology

基　　金：贵州省教育厅自然科学研究招标项目基金(黔教合KY字2015375)资助

摘　　要：齐墩果酸（oleanolic acid,OA）是一种广泛存在于自然界中的五环三萜类天然化合物,已有研究表明OA具有抗肺癌活性,但是具体机制尚未完全阐释清楚。本研究旨在证实OA是否能够诱导非小细胞肺癌A549细胞凋亡,并探索该效应是否与线粒体凋亡通路相关。我们将不同浓度的OA作用于细胞后,使用MTT法检测A549细胞的生长情况。采用AV/PI法进行染色后,经流式细胞仪检测其凋亡率,同时使用Hoechst 33342染色并使用荧光显微镜观察并拍照;Westen-blotting检测OA对A549细胞的线粒体凋亡通路相关蛋白相对表达水平的影响。MTT结果显示OA能够抑制A549细胞的生长,并呈时间剂量依赖性。AV/PI法染色结果显示OA能够诱导A549细胞凋亡,亦呈时间剂量依赖性。Hoechst 33342法染色结果相似。Westen-blotting结果表明OA能够上调A549细胞线粒体凋亡通路相关蛋白Bax,t Bid,Cleaved caspase 3的相对表达水平,同时下调该通路Bcl-xl,Bcl-2的相对表达水平。以上所有的结果表明OA能够显著抑制肺癌A549细胞的生长并通过调节线粒体凋亡通路相关蛋白的表达水平而诱导其凋亡。Oleanolic Acid（OA, 3β-hydroxyolean-12-en-28-oic acid） as a natural triterpenoid is ubiquitous in many dietary and medicinal plants. Experimental and clinical studies reported that it has antitumor activity,unfortunately, the underlying molecular mechanisms have not yet been fully explained, studies on anti-cancer effect of oleanolic acid are still insufficient. In this study, we focus on whether the oleanolic acid can induce the apoptosis of non-small cells lung cancer A549 cell line by mediating the mitochondrial apoptotic pathway. We detected the proliferation of A549 cells by MTT assay after treated with different concentrations of oleanolic acid.The apoptotic rate was investigated by flow cytometry after stained with AV/PI. Simultaneously, when staining with Hoechst 33342, we could intuitively observe the apoptotic cells and photographed by using fluorescence microscope. The results of MTT showed that oleanolic acid could inhibit the A549 cells in a dose-dependent and time-dependent manner. Data of flow cytometry demonstrated that oleanolic acid induced the apoptosis of A549 cells in a dose-dependent and time-dependent manner similarly. The data of staining with Hoechst 33342 were consistent with data of flow cytometry. Additionally, western blotting analysis suggested that Oleanolic acid could up-regulate the expressions of relative proteins in mitochondrial apoptotic pathway like Bax, t Bid, Cleaved caspase 3 and down-regulated the expressions of Bcl-xl, Bcl-2. All evidence suggested that OA could inhibit the proliferation of A549 cell line and induce apoptosis by regulating the expression of relative mitochondrial apoptotic molecular.

关 键 词：齐墩果酸 A549细胞 凋亡 线粒体通路 

分 类 号：R285[医药卫生—中药学]