壁虎醇提物对人食管鳞癌细胞EC9706的作用和体内抗肿瘤活性  

Effect of gecko alcohol extract on human esophageal squamous carcinoma cell line EC9706 and anti-tumor activity in vivo

作　　者：王晓兰[1] 王淑英[1] 王建刚[1] 

机构地区：[1]河南科技大学医学院,河南洛阳471003

出　　处：《中国中药杂志》2010年第16期2175-2179,共5页China Journal of Chinese Materia Medica

基　　金：河南省医学科技攻关计划项目(200903109)

摘　　要：目的：观察壁虎醇提物体内外抗肿瘤作用。方法：体外实验采用四噻唑蓝法（MTT）检测壁虎醇提物对人食管鳞癌细胞EC9706的增殖抑制作用;采用Hoechst33258荧光染色观察细胞凋亡的形态改变;采用TUNEL法检测细胞凋亡率;采用SP免疫组化法检测细胞内凋亡蛋白Bax和Bc l-2的表达。体内实验以小鼠S180肉瘤为模型,观察壁虎醇提物对肿瘤生长的抑制作用。结果：6-8 g.L-1壁虎醇提物作用于细胞24,48,72 h后能够明显抑制EC9706食管鳞癌细胞株的生长（P（0.01）,抑制作用呈剂量和时间依赖性;Hoechst33258荧光染色可见凋亡细胞;6,7 g.L-1壁虎醇提物处理组细胞凋亡率高于阴性对照组（P〈0.05,P〈0.01）,凋亡率分别为（12.73±3.84）%,（19.80±2.32）%;免疫组化结果显示细胞内Bc l-2蛋白无明显变化,而Bax蛋白表达升高（P（0.01）,Bax/Bc l-2升高。不同剂量的壁虎醇提物（0.6,1.2,2.4 g.kg-1）均可抑制S180肉瘤在昆明小鼠体内生长,抑瘤率分别为44.88%,63.94%,69.53%。结论：壁虎醇提物体内体外具有抗肿瘤作用,体外诱导EC9706细胞凋亡的作用机制可能与凋亡相关蛋白Bax/Bc l-2升高有关。Objective： To observe the anti-tumor activity of gecko alcohol extract in vitro and in vivo.Method： in vitro,the inhibitory effect of gecko alcohol extract on proliferation of human esophageal carcinoma EC9706 cells was measured by MTT colorimetric assay.Morphological change of apoptotic cells was observed by Hoechst33258 fluorescence staining and apoptosis induced by gecko alcohol extract was evaluated in TUNEL assay.The expression of apoptosis protein Bax and Bcl-2 in EC9706 cells was investigated by immunohistochemistry.in vivo,the inhibitory effect of gecko alcohol extract on tumor growth was examined on S180 sarcoma model.Result： After gecko alcohol extract（6-8 g·L-1） treatment for 24,48 and 72 h separately,EC9706 cell proliferation was significantly inhibited in both dose-and time-dependent manner（P 0.01）.Cell apoptosis in gecko alcohol extract-treated group（6,7 g·L-1） was significantly higher than that in control group[（12.73±3.84） %,（9.80±2.32） % vs（5.87±2.54） %,P 0.05,P0.01].Although the level of Bcl-2 did not change significantly,the expression of Bax was rembarkably up-regulated in gecko alcohol extract treated group.Gecko alcohol extract inhibited the growth of S180 sarcoma in Kun-ming mice at all doses（0.6,1.2,2.4 g·kg-1） of administration.The inhibitory rate was 44.88%,63.94% and 69.53% respectively.Conclusion： From tumor inhibitory test,gecko alcohol extract shows significantly inhibitory effect in vitro and in vivo.The increased Bax/Bcl-2 ratio is mechanistically linked to the gecko alcohol extract-induced tumor cell apoptosis.

关 键 词：壁虎醇提物 MTT法 食管癌细胞株 TUNEL法 凋亡蛋白 S180荷瘤小鼠 

分 类 号：R285[医药卫生—中药学]