牵牛子酒提取物对Lewis肺癌的抗肿瘤和抗转移机制研究  

Study on anti-tumor and anti-metastasis mechanism of alcohol extracts from Pharbitidis Semen against Lewis lung cancer

作　　者：李佳桓[1] 杜钢军[1] 刘伟杰[1] 刘迎辉[1] 赵贝[1] 李红[1] 

机构地区：[1]河南大学药学院药物研究所,河南开封475001

出　　处：《中国中药杂志》2014年第5期879-884,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81173094);河南省科技厅重点攻关项目(112101310308);河南大学省部共建项目(SBGJ090704);河南省高校青年骨干教师项目(2010GGJS-025)

摘　　要：目的:观察牵牛子酒提取物对Lewis肺癌生长和转移的影响,研究其抗肿瘤机制。方法:体外采用MTT法和划痕实验检测牵牛子酒提取物对Lewis肺癌细胞生长和迁移的影响,吖啶橙染色检测细胞自噬,荧光黄传输检测细胞间连接通讯,Western blotting检测细胞中水通道蛋白1(AQP1)的表达。体内建立小鼠Lewis肺癌皮下移植模型和实验性肺转移模型评价牵牛子酒提取物的抗肿瘤和抗转移作用,电化学发光法检测荷瘤小鼠血清癌胚抗原(CEA)和β2微球蛋白(β2-MG),免疫组化法检测肺转移小鼠肺脏AQP1和缝隙连接蛋白Cx43表达。结果:牵牛子酒提取物对体外培养的Lewis肺癌细胞呈剂量依赖性生长抑制,明显阻止细胞迁移,降低细胞AQP1蛋白,促进细胞间连接通讯,减少癌细胞无血清自噬。体内实验,与未治疗的模型小鼠比较,牵牛子酒提取物呈剂量依赖性减缓肿瘤生长、阻止肿瘤转移、延长荷瘤小鼠存活时间,同时,牵牛子酒提取物也降低荷瘤小鼠血清CEA和β2-MG水平,能增强荷瘤肺组织间隙连接蛋白Cx43的免疫组化染色深度,减弱水通道蛋白AQP1的阳性染色密度。结论:牵牛子酒提取物能够阻止Lewis肺癌生长和转移,其机制可能与增强细胞间连接通讯和下调细胞水通道AQP1有关。Objective： To observe the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer, and study its anti-tumor mechanism. Method： In vitro, M3T assay and scratch assay were adopted to detect the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer cells. The cell autophagy was detected by the acridine orange staining. The gap-junction intercellular communication ~ GJIC ） was investigated by the fluorescent yellow transfer. The expression of aquaporin 1 （ AQP1 ） was analyzed by the Western blotting. In vivo, the subcutaneous implant model and the experimental pulmonary metastasis model of Lewis lung cancer in mice were established to evaluate the anti-tumor and anti-metastasis effects of alcohol extract from Pharbitidis Semen. The serum carcinoembryonic antigen （CEA） and/32 microglobulin （/32-MG） of mice bearing lewis lung cancer were detected by the electrochemiluminesence immunoassay. The expressions of lung AQP1 and Con- nexin 43 （Cx43） were examined by the immunohistochemical method. Result： In vitro, alcohol extracts from Pharbitidis Semen inhibi- ted the cell proliferation in a dose-dependent matter, significantly prevented the cell migration, down-regulated AQP1 proteins of cells, promoted GJIC, and decreased the serum-free autophagy of tumor cells. In vivo, compared with untreated model mice, alcohol extracts from Pharbitidis Semen inhibited the tumor growth in a dose-dependent matter, prevented the tumor metastasis and prolonged the life span of mice bearing Lewis lung cancer, while decreasing serum CEA and/32-MG of mice bearing Lewis lung cancer, enhancing the immumohistochemical staining intensity of Cx43 and weakening aquaporins AQP1 positive intensity. Conclusion： Alcohol extracts from Pharbitidis Semen could prevent the proliferation and metastasis in Lewis lung cancer cells. Its mechanism may be related to the promotion of GJIC and the down-regulation of AQP1.

关 键 词：牵牛子酒提取物 LEWIS肺癌 细胞间连接通讯 水通道蛋白1 

分 类 号：R284.2[医药卫生—中药学]