Effect of Sesamin on Proliferation and Apoptosis of Pulmonary Adenocarcinoma Cell
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摘要: 目的 探讨芝麻素对人肺腺癌A549细胞株的抗增殖和诱导凋亡作用及其机制，为开发抗肿瘤中药提供实验和理论依据。方法 应用MTT法检测芝麻素对A549细胞株的抗增殖及细胞毒性作用；应用倒置显微镜、HE染色法来观察肿瘤细胞的形态学变化；应用免疫细胞化学技术检测Caspase-3、Caspase-8、Caspase-9的表达变化；AnnexinV/PI双染色法分析细胞凋亡率及测定细胞Bcl-2及Bax表达水平。结果 芝麻素对A549细胞株均有明显抑制作用，且呈现出浓度和时间依赖性；倒置显微镜观察发现实验组细胞体积变小变圆，细胞间连接疏松，贴壁能力减弱；HE染色结果发现实验组细胞质脱水浓缩，伊红染色增强等典型的细胞形态学变化，免疫细胞化学检测发现实验组细胞Caspase-3、Caspase-8和Caspase-9的表达增加与对照组相比较差异有统计学意义(P<0.01)；流式细胞仪分析结果示芝麻素40 μg/m1作用48 h后，细胞主要阻滞在S期；Bcl-2蛋白的表达降低，Bax 蛋白的表达升高。结论 芝麻素对人肺腺癌A549细胞株具有抑制增殖作用，并呈浓度和时间依赖性佳；芝麻素诱导凋亡作用机制可能与细胞周期发生S期阻滞有关；芝麻素由死亡受体通路和线粒体通路共同完成凋亡的启动和执行。
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关键词:
- 芝麻素 /
- 人肺腺癌A549细胞 /
- 细胞凋亡 /
- 增殖
Abstract: Objective To investigate the effect and mechanism of sesamin on antiproliferation and apoptosis induction of human lung adenocarcinoma cell line A-549 in vitro, and provide a theory basis for traditional medicine treatment in antineoplastic therapy. Methods Human lung adenocarcinoma cell line A549 was cultivated in vitro. Then the effect of antiproliferation and cytotoxicity of sesamin on human lung adenocarcinoma cell line A549 was analyzed by MTT; Morphological changes were observed by inverted microscope and HE staining; Flow cytometry was used to investigate the expression of Bcl-2 and Bax; The apoptosis rate was determined by AnnexinV/PI staining; The changes of apoptosis protein kinase such as Caspase-3, Caspase-8, Caspase-9 were detected by immunohistochemistry. Results MTT assay showed that sesamin had signifi cant inhibition on the growth of A549 cell line in dose- and time-dependent manner; It was observed by inverted microscope the volume of the cell A549 was decreased and got round, nuclear condensed chromatin, the junction between cell was loosed, adherence ability was decreased. A typical cell morphology changes such as the cytoplasm dehydrated and concentrated was showed by HE staining and enhanced feosin staining; Immunohistochemical assay showed that the expression of Caspase-3, 8, 9 was increased compared with those in control group, it had a signifi cantly difference (P<0.05); Flow cytometry showed a cell apoptosis peak after treated by 40 μg/ml sesamin for 48 h, and the number of cell was increased in S phase and decreased in G0/G1 phase, and most cell stay in S phase. The expression of Bcl-2 was decreased, while the expression of Bax was increased. Conclusion Sesamin can signifi eantly inhibit the growth of A549 cells and the effect is dose-dependent and time-dependent. Sesamin could arrest the A549 cells in the S phase. Sesamin induced apoptosis via two main pathways: the mitochondrial pathway and the death receptor-mediated pathway. Sesamin has a signifi cant effect on anti-tumor. It will be hopefully exploited as a new anti-tumor or tonic.-
Key words:
- Sesamin /
- Human lung cancer cell line A549 /
- Apoptosis /
- Proliferation
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