眼镜蛇毒神经生长因子诱导人鼻咽癌CNE-2细胞凋亡作用的研究  

Study on the apoptosis induction of Guangxi cobra venom NGF in human nasopharyngeal carcinoma CNE-2 cells

作　　者：韦世秀[1] 刘成军[2] 李牡艳[1] 宋慧[3] 李佳荃[1] 

机构地区：[1]广西医科大学医学科学实验中心,广西南宁530021 [2]广西壮族自治区江滨医院药剂科,广西南宁530021 [3]广西医科大学化学教研室,广西南宁530021

出　　处：《中国医院药学杂志》2008年第22期1909-1912,共4页Chinese Journal of Hospital Pharmacy

基　　金：广西科技厅科学基金项目(编号:0542043)

摘　　要：目的:研究广西眼镜蛇毒神经生长因子(Nerve growth factor,NGF)对人鼻咽癌CNE-2细胞增殖的抑制作用和诱导细胞凋亡。方法:应用三磷酸腺苷-生物荧光肿瘤化疗药物敏感试验法(ATP-TCA法)测定不同质量浓度(0.0125,0.025,0.05,0.1,0.2g.L-1)NGF处理24,48,72,96h对人鼻咽癌CNE-2生长的抑制率;Hoechst33342荧光染色观察凋亡细胞的形态;琼脂糖凝胶电泳测定DNA断裂状况;流式细胞术检测细胞周期变化和细胞凋亡率。结果:不同浓度的NGF能抑制CNE-2细胞增殖,并呈浓度-时间效应关系,作用24,48,72,96h的半数抑制浓度(IC50)分别为0.213,0.095,0.048,0.033g.L-1;经NGF(0.1g.L-1)处理细胞48h后,荧光染色呈现典型的细胞凋亡形态学特征;DNA凝胶电泳可见凋亡细胞特有的DNA片段;0.05,0.1,0.2g.L-1NGF作用48h后,CNE-2细胞被阻滞于G0/G1期,细胞凋亡率分别为(28.2±2.0)%、(39.9±1.9)%、(50.3±1.3)%。结论:NGF通过诱导人鼻咽癌CNE-2细胞凋亡而产生抗鼻咽癌活性。OBJECTIVE To study the proliferation inhibition and apoptosis induction of Guangxi cobra venom NOF（Nerve growth factor） in human nasopharyngeal carcinoma CNE-2 cells. METHODS To detect the inhibition ratio of the human nasopharyngeal carcinoma CNE-2 cells by Adenosine. The adenosine triphosphate （ATP）-based tumor chemosensitivity assay （ATP-TCA） in different time （24h, 48h, 72h, 96h later） treated by NGF in different dosages （0. 0125,0. 025,0. 05,0. 1,0. 2 g · L ^-1）. Morphology of the apoptosis cells was observed by Hoechst 33342 fluorescein stain. DNA ladder straps were detected by agarose gel electrophoresis. Changes of the cell cycles and the ratio of the apoptosis were detected by flow eytometry. RESULTS The experiment showed that NGF could obviously inhibit the proliferation of CNE2 cells in a concentration and times dependent manner, the IC50 values of NGF at 24 h,48 h,72 h,96 h were 0. 213 g· L ^-1,0. 095 g · L ^-1,0. 04 g · L ^-1 and 0. 033 g · L ^-1 ;Fluorescein stain presented typical morphology of apoptosis after 48 h treated by NGF （0. 1 g · L ^-1 ）. DNA ladder straps were shown in gel electrophoresis. CNE-2 cells were blocked in G0/G1 stage treated by NGF （0. 05,0. 1,0.2 g · L ^-1） after 48h, and the ratio of the apoptosis were（28. 2 ± 2.0） %, （39. 9 ± 1.9） % and（50. 3 ± 1.3） %. CONCLUSION NGF have anti-human nasopharyngeal carcinoma effect by induction apoptosis of the CNE-2 cells.

关 键 词：广西眼镜蛇毒 神经生长因子 鼻咽癌 CNE-2细胞 细胞凋亡 

分 类 号：R285[医药卫生—中药学] R730.5[医药卫生—中医学]