鹅不食草提取物对人鼻咽癌细胞CNE-1增殖抑制和凋亡诱导作用  

Proliferation inhibition and apoptosis induction of Centipeda minima extracts on human nasopharyngeal carcinoma cell CNE-1

作　　者：郭育卿[1,2] 王文强[2] 陈志安[2] 陈四保[1,2] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]香港理工大学中药药学及分子药理学国家重点实验室培育基地,深圳518057

出　　处：《生物加工过程》2013年第3期65-70,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金(30873150);"香港中药材标准"项目

摘　　要：考察鹅不食草提取物对人高分化鼻咽癌细胞CNE-1增殖的抑制作用,初步探讨其抗鼻咽癌分子机制。95%乙醇提取鹅不食草全草,噻唑蓝(MTT)法观察不同浓度提取物在不同时间对人高分化鼻咽癌细胞CNE-1增殖的抑制作用,Hoechst 33258荧光染色观察CNE-1凋亡情况及形态学变化,Western Blot检测CNE-1细胞内抗凋亡蛋白Bcl-2和促凋亡蛋白Bax的表达。结果表明:鹅不食草醇提物能显著抑制CNE-1增殖(P<0.01),并呈现明显的时间-剂量依赖性,其中诱导48和72 h的IC50分别为30.0和25.0μg/mL。阳性对照药顺铂的IC50为0.79μg/mL。Hoechst 33258荧光染色观察发现给药组CNE-1细胞出现不同程度细胞核变圆变亮,核染色质浓缩,产生核小体等明显的凋亡特征。Bcl-2蛋白相对表达量随醇提物浓度的增大而下降,Bax蛋白相对表达量随醇提物浓度的增大而上升,二者与对照组相比均有显著性差异(P<0.01)。鹅不食草醇提物对体外人高分化鼻咽癌细胞CNE-1具有明显的增殖抑制和凋亡诱导作用,其分子作用机制可能与Bcl-2蛋白表达下调、Bax蛋白表达上调有关。To investigate proliferation inhibition and apoptosis induction of Centipeda minima extracts on human nasopharyngeal carcinoma cell CNE-1, and explore its potential mechanism. Whole herb of Centipeda minima was extracted with 95% ethanol. After treated with various concentrations of extracts for 48 h, and 72 h, the cell growth of CNE-1 was evaluated by MTT assay. Morphologic changes of extract- treated CNE-1 cells were observed by light microscopy. Apoptotic cells were visualized by Hoechst 33258 staining by inversion fluorescence microscopy. Expressions of Bax and Bcl-2 protein were detected by Western Blotting. Etharol extract of Centipeda minima showed significant anti-proliferative activities（ P 〈 0. 01 ） and apoptosis induction on CNE-1 in a time-concentration dependent manner. The IC50 values were 30. 0 μg/mL at 48 h ,and 25.0 μg/mL at 72 h incubation,respectively. The cisplatin ICs0value was 0. 79 μg/mL after incubating CNE-1 for 72 h. Typical condensed chromatins and fragmented nuclear bodies were observed in apoptotic cells after Ho 33258 staining. Western Blotting test showed that Bax protein was up-regulated while Bcl-2 protein was down-regulated after treated with the extract for 48 h, and had significant variation compared with the Control（ P 〈 0.01 ）. Centipeda minima ethanol extract had significant anti-proliferation and apoptosis induction effects on CNE-1 in vitro, and its mechanism was related to up-regulation of Bax protein and down-regulation of Bcl-2 protein.

关 键 词：凋亡 鹅不食草 鼻咽癌 CNE-1 增殖 

分 类 号：R285.5[医药卫生—中药学]