芹菜籽提取物通过使BGC - 823人胃癌细胞系的细胞周期停滞于S期诱导细胞凋亡的分子机制。
Molecular mechanisms of celery seed extract induced apoptosis via s phase cell cycle arrest in the BGC-823 human stomach cancer cell line.
作者信息
Gao Lin-Lin, Feng Lei, Yao Shu-Tong, Jiao Peng, Qin Shu-Cun, Zhang Wei, Zhang Ya-Bin, Li Fu-Rong
机构信息
Department of Pathology and Pathophysiology, Taishan Medical University, Taian, China.
出版信息
Asian Pac J Cancer Prev. 2011;12(10):2601-6.
BACKGROUND
Mechanisms of apoptosis in tumor cells is an important field of tumor therapy and cancer molecular biology. Loss of cell cycle control, leading to uncontrolled proliferation, is common in cancer. Therefore, the identification of potent and selective cyclin dependent kinase inhibitors is a priority for anti-cancer drug discovery. There are at least two major apoptotic pathways, initiated by caspase-8 and caspase-9, respectively, which can activate caspase cascades. Apoptosis triggered by activation of the mitochondrial-dependent caspase pathway represents the main programmed cell death mechanism. This is activated by various intracellular stresses that induce permeabilization of the mitochondrial membrane. Anti-tumor effects of celery seed extract (CSE) and related mechanisms regarding apoptosis were here investigated in human gastric cancer BGC-823 cells.
METHODS
CSE was produced by supercritical fluid extraction. Cell viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide (MTT) assay and apoptosis by flow cytometry using Annexin/PI staining and DAPI staining and a laser scanning confocal microscope (LSCM). Cell cycling was evaluated using PI staining with flow cytometry and expression of cell cycle and apoptosis-related proteins cyclin A, CDK2, bcl-2 and bax was assessed by immunohistochemical staining.
RESULTS
CSE had an anti-proliferation effect on human gastric cancer BGC-823 cells in a dose- and time-dependent manner. After treatment, the apoptotic rate significantly increased, with morphological changes typical of apoptosis observed with LSCM by DAPI staining. Cell cycle and apoptosis related proteins, such as cyclin A, CDK2 and bcl-2 were all down-regulated, whereas bax was up-regulated.
CONCLUSIONS
The molecular determinants of inhibition of cell proliferation as well as apoptosis of CSE may be associated with cycle arrest in the S phase.
背景
肿瘤细胞凋亡机制是肿瘤治疗和癌症分子生物学的一个重要领域。细胞周期控制的丧失导致不受控制的增殖，这在癌症中很常见。因此，鉴定强效和选择性的细胞周期蛋白依赖性激酶抑制剂是抗癌药物发现的首要任务。至少有两条主要的凋亡途径，分别由半胱天冬酶 - 8和半胱天冬酶 - 9启动，它们可以激活半胱天冬酶级联反应。由线粒体依赖性半胱天冬酶途径激活引发的凋亡代表了主要的程序性细胞死亡机制。这是由各种诱导线粒体膜通透性的细胞内应激激活的。本文研究了芹菜籽提取物（CSE）对人胃癌BGC - 823细胞的抗肿瘤作用及其凋亡相关机制。
方法
通过超临界流体萃取制备CSE。采用3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基溴化四氮唑（MTT）法分析细胞活力，使用膜联蛋白/碘化丙啶染色、4',6-二脒基-2-苯基吲哚（DAPI）染色和激光扫描共聚焦显微镜（LSCM）通过流式细胞术分析凋亡。使用PI染色通过流式细胞术评估细胞周期，并通过免疫组织化学染色评估细胞周期和凋亡相关蛋白细胞周期蛋白A、细胞周期蛋白依赖性激酶2（CDK2）、bcl-2和bax的表达。
结果
CSE对人胃癌BGC - 823细胞具有剂量和时间依赖性的抗增殖作用。处理后，凋亡率显著增加，通过DAPI染色在LSCM下观察到典型的凋亡形态变化。细胞周期和凋亡相关蛋白，如细胞周期蛋白A、CDK2和bcl-2均下调，而bax上调。
结论
CSE抑制细胞增殖以及凋亡的分子决定因素可能与S期的细胞周期停滞有关。
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