眼镜蛇毒L-氨基酸氧化酶诱导人胃癌细胞MGC-803细胞凋亡的实验研究  

Evidence of caspases-mediated human stomach carcinoma MGC-803 cells apoptosis induced by L-amino acid oxidase from Naja atra venom

作　　者：齐元麟[1] 张明芳[1,2] 赵蓉[1] 林建银[2] 

机构地区：[1]福建医科大学基础医学院分子医学研究中心消化道恶性肿瘤教育部重点实验室 [2]福建医科大学生理学与病理生理学系

出　　处：《中国临床药理学与治疗学》2012年第2期121-125,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金(30940062);福建医科大学青年科学基金(FJGXQ04011);福建省教育厅科技重点项目(JA10130)

摘　　要：目的:研究眼镜蛇毒L-氨基酸氧化酶(NA-LAAO)对人胃癌细胞MGC-803的细胞毒性、诱导凋亡作用及可能机制。方法:采用CCK-8法测定细胞毒性;采用DNA倍体分析和An-nexin V/碘化丙啶染色测定细胞凋亡;采用发色底物法测定Caspase-3酶活性;采用Western-blot方法测定聚二磷酸腺苷核糖多聚酸-1(PARP-1)切割。结果:NA-LAAO可抑制MGC-803细胞增殖,6、12、24h的IC50分别为(2.48±0.41)、(1.74±0.27)和(0.83±0.19)mg/L;NA-LAAO可使细胞DNA分布出现亚二倍体凋亡峰,并可促使细胞膜磷脂酰丝氨酸外翻;NA-LAAO可激活Caspase-3并可促使其底物PARP-1降解。结论:NA-LAAO可能通过激活Caspase-3这一分子机制而诱导细胞凋亡,从而抑制肿瘤细胞增殖。AIM. To investigate the effects and the molecular mechanisms of Naja atra L-a- mino acid oxidase （NA-LAAO） on the proliferation and cell apoptosis of human stomach cancer MGC-803 cells. METHODS.. MGC-803 cells were exposed to different concentrations of NA- LAAO and the cytotoxicity was measured using a CCK-8 kit. The apoptosis-inducing activities of NA-LAAO were examined by propidium iodide staining and Annexin V/PI assay in flow cytometry. The enzyme activity of caspase-3 was measured by a colorimetric assay. The cleavage of PARP-1 was analyzed by Western-blot. RESULTS. NA-LAAO exhibited cytotoxicity on MGC-803 in a dose- and time-dependent manner. The half maximal inhibitory concentration （IC50） values of NA-LAAO were （2.48±0.41）, （1.74 ±0.27） and （0.83±0.19） mg/L in MGC-803 cells at 6 h, 12 h and 24 h, respectively. NALAAO induced apoptosis accompanied DNA hy- podiploidy, phosphatidylserine exposure, activation of caspase-3, and PARP cleavage. CONCLUSION： NA-LAAO inhibited cell proliferation by inducing a caspase-mediated cell apoptosis.

关 键 词：蛇毒 中华眼镜蛇 L-氨基酸氧化酶 细胞增殖 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤] R73-3[医药卫生—临床医学]