中国药理学通报2011，Vol.27Issue(12)：1660-1665,6.DOI:10.3969/j.issn.1001-1978.2011.12.009
眼镜蛇毒L-氨基酸氧化酶对人膀胱癌ECV304细胞增殖和细胞周期的影响及机制
Effects of Naja atra L-amino acid oxidase on the proliferation and cell cycle of human bladder carcinoma ECV304 cells
摘要
Abstract
Aim To investigate the effects and the molecular mechanisms of Naja atra L-amino acid oxidase ( NA-LAAO ) on the proliferation and cell cycle of human bladder carcinoma ECV304 cells. Methods ECV304 cells were exposed to different concentrations of NA-LAAO and the cytotoxicity was measured by CCK-8 method. The effects of NA-LAAO on cell cycle were examined by a propidium iodide staining in flow cytometry. The mRNA or protein expressions of cell cycle regulators were determined by real-time PCR or Western blot, respectively. Results NA-LAAO exhibited cytotoxicity in ECV304 in a dose- and time-dependent manner. The half maximal inhibitory concentration ( IC50 ) values of NA-LAAO were ( 1. 54 ± 0. 23 ),( 1. 09 ± 0. 15 ) and ( 0. 48 ± 0. 14 ) mg · L-1 in ECV304 cells at 6 h, 12 h and 24 h, respectively. At concentrations lower than 0. 625 mg · L-1, NA-LAAO did not kill the ECV304 cells but inhibited cellproliferation. Four days ' exposure to NA-LAAO of 0. 156 mg · L-1 or two days' exposure to NA-LAAO of 0. 313 mg · L-1 caused a significant decrease of cell growth. Analysis of ECV304 cell cycle by flow cytometry method indicated that NA-LAAO could increase the G1-phase cells and block the cell cycle in G1-phase. Real-time PCR analysis indicated NA-LAAO up-regulated the mRNA expression of p21, pi 6, p27 whereas down-regulated the expression of cyclin A2, cyclin Bl, cyclin Dl, cyclin El, CDK2, CDK6, Survivin and Skp2. Western blot analysis indicated NA-LAAO up-regulated p21 and down-regulated cyclin A, CDK2 and CDK6. These effects might contribute to the blockade of cell cycle. Conclusion NA-LAAO inhibits cell proliferation by blocking the cell cycle in G1 -phase.关键词
蛇毒/中华眼镜蛇/L-氨基酸氧化酶/细胞增殖/细胞周期/膀胱癌Key words
snake venoms/ Naja atra/ L-amino acid oxidase/ cell proliferation/ cell cycle/ urinary bladder carcinoma分类
医药卫生引用本文复制引用
齐元麟,张明芳,赵蓉,张旭,吴玲珊,林建银..眼镜蛇毒L-氨基酸氧化酶对人膀胱癌ECV304细胞增殖和细胞周期的影响及机制[J].中国药理学通报,2011,27(12):1660-1665,6.基金项目
国家自然科学基金资助项目(No 30371747) （No 30371747）
福建省教育厅科技重点资助项目(No JA10130) （No JA10130）
福建医科大学青年科学基金资助项目(No FJGXQ04011) （No FJGXQ04011）
福建医科大学大学生创新性实验计划资助项目(No C10050) （No C10050）
