| 李发凯,张 芳,陆 远,王 媛,李 昂,吴健雄,任新玲.紫草素促进肺癌A549细胞凋亡[J].现代生物医学进展英文版,2018,(9):1611-1615. |
| 紫草素促进肺癌A549细胞凋亡 |
| Shikonin Promotes the Apoptosis of A549 Human Lung Cancer Cells |
| Received:October 18, 2017 Revised:November 12, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.09.003 |
| 中文关键词: 紫草素 肺癌 凋亡 |
| 英文关键词: Shikonin Lung cancer Apoptosis |
| 基金项目:国家自然科学基金项目（81172222） |
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| 中文摘要: |
| 摘要 目的：探讨紫草素对A549人肺癌细胞凋亡的影响和可能的作用机制。方法：采用不同浓度的紫草素对体外培养的A549人肺癌细胞进行干预，CCK-8法和流式细胞术分别检测紫草素对A549细胞增殖和凋亡的影响，Western blot观察凋亡相关蛋白（Bcl-2和Bax）表达水平的变化，激光共聚焦显微镜检测紫草素处理12 h并用JC-1染色的A549细胞线粒体膜电位改变。结果：CCK-8分析显示， 0.5 μM、1 μM、2 μM、4 μM和6 μM实验组A549细胞相对存活率分别为(83.71±1.02)%、(57.47±2.78)%、(27.39±1.96)%、(16.96±1.47)%和(14.72±1.93)%，与对照组相比实验组A549细胞相对存活率明显降低；流式细胞术结果表明，1 μM、2 μM、4 μM实验组A549细胞的凋亡率分别为(13.80±1.76)%、(40.90±3.48)%和(78.80±2.52)%，与对照组相比紫草素呈剂量依赖型促进A549细胞凋亡； Western blot 结果证实，紫草素能降低A549细胞中Bcl-2蛋白的表达量，而升高Bax蛋白的水平；激光共聚焦显微镜扫描结果显示紫草素能降低A549细胞的线粒体膜电位，呈剂量依赖型。结论：紫草素能显著促进A549细胞凋亡，其机制可能与下调抗凋亡蛋白Bcl-2的表达和上调促凋亡蛋白Bax的表达有关。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of shikonin on apoptosis of A549 human lung cancer cells, as well as the mechanism. Methods: The A549 cells were treated with different concentrations of shikonin in vitro. The cell viability was assessed by CCK-8 Assay, and the cell apoptosis was determined by flow cytometry. The expression of apoptosis associated proteins(Bcl-2 and Bax), was observed by Western blot. The MMP（mitochondrial membrane potential） of A549 cells was detected by Laser scanning Confocal Microscopy(LSCM) using JC-1 probes 12 h after shikonin treatment. Results: After exposure to various concentrations of shikonin(0.5μM, 1 μM, 2 μM, 4 μM and 6 μM), as the results of CCK-8 showed, the survival rates of A549 cells were (83.71±1.02)%, (57.47±2.78)%, (27.39±1.96)%, (16.96±1.47)% and (14.72±1.93)%, respectively, which were obviously lower than those in the control group. Further study on shikonin showed that it induced apoptosis of A549 cells in a dose-dependent manner; the apoptosis rates in shikonin-treated groups(1 μM, 2 μM, 4 μM ) were (13.80±1.76)%, (40.90±3.48)% and (78.80±2.52)%, respectively, compared to those in the control group. In addition, a western blot analysis indicated that shikonin decreased the expression of protein Bcl-2, but increased the expression of pro- tein Bax. Moreover, shikonin depolarized the MMP of A549 cells in a dose-dependent manner too. Conclusion: Our results indicated that shikonin significantly induced apoptosis of A549 cells, the mechanism may be related to the down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of apoptotic protein Bax. |
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