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尾静脉注射EMT-6细胞株建立原位肺癌动物模型的研究
Establishment of an orthotopic lung cancer model by injection of EMT-6 cells
魏诗航,韦世元,施雪旎,彭旭,何学令,刘艳,尹海林
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作者单位:四川大学生命科学学院;四川大学生命科学学院;四川大学生命科学学院;四川大学实验动物中心;四川大学实验动物中心;四川大学实验动物中心;四川大学实验动物中心;
中文关键字:EMT-6;不同浓度;转移性肺癌动物模型
英文关键字:EMT-6; different concentrations;animal model of metastasis lung cancer;
中文摘要: 目的 通过尾静脉注射EMT-6癌细胞,建立一种临床特征明显、稳定可重复的转移性肺癌BALB/c小鼠模型。 方法 取对数生长期的EMT-6乳腺癌细胞,用0.25%胰蛋白酶消化,再用PBS调整细胞浓度为1*106/ml(高)、5*105/ml(中)和1*105/ml(低)三个浓度,尾静脉注0.2mlEMT-6癌细胞悬液,对不同浓度模型的临床症状、成瘤时间、生存期、成瘤程度和病理学特征等生物学特征和评价指标进行研究,筛选出效果好的模型,并对该模型的重复性和稳定性通过三次重复实验加以确定,成功建立EMT-6细胞转移性肺癌BALB/c小鼠模型。 结果 EMT-6细胞注射后解剖实验动物发现,高浓度(1*106/ml)动物模型7d肺部表面开始出现肿瘤灶,18d内所有动物死亡;中浓度(5*105/ml)动物模型7d肺部表面开始出现肿瘤灶,28d内所有动物死亡;低浓度(1*105/ml)动物模型14d肺部表面开始出现肿瘤灶,21d所有动物肺部表面均有肿瘤灶,荷瘤动物28d开始出现死亡,35天瘤灶数量达到顶峰(平均12-15个/只)且瘤体积变大,42d内全部死亡。相比之下低浓度模型成瘤时间到动物全部死亡时间,为实验研究留有4周的处理、观察、评价的窗口期,其病理组织学符合肺癌的组织学特征,临床症状指标明显,三次测试模型重复性和稳定性好,是适合转移性肺癌研究的理想模型。
英文摘要: Objective: To establish a metastasis lung cancer model, characteristics is consistent with clinical, stable and repeatable, in BALB/c mice by intravenous injection of EMT-6 breast-tumor cells. Methods: Harvest the logarithmic growth phase cells of EMT-6 murine mammary carcinoma by 0.25% trypsin digestion, and equilibrated at 3 density of 1*106 cells per 1ml(high), 5*105 cells per 1ml(medium) and 1*105 cells per 1ml(low) in phosphate-buffered saline(PBS). And then intravenously inject 0.2ml EMT-6 tumor cells. Do research on evaluation indicators and biological characteristics such as clinic appearance, the time of tumor formation, survival time, the tumor growth and pathological characteristics of metastasis lung models with different concentrations. And then screen the best metastasis lung cancer model. Finally determine the repeatability and stability of the model by 3 repeated experiment, successfully established a metastasis lung cancer model in BALB/c mice. Results: Anatomized BALB/c mice after EMT-6 tumor cells injection, it showed that in the group of high concentration (1*106/ml), tumor nodes on lung surface began to appear at 7d and all animals died in 18d; in the group of medium concentration(5*105/ml), tumor nodes on lung surface began to appear at 7d and all animals died in 28d; in the group of low concentration(1*105/ml), tumor nodes on lung surface appeared partially at 14d, all mice lung surface had tumors at 21d, the number of tumors reach the highest level(an average of 12-15/mouse) and volume of tumors became much bigger at 35d, mice began to die at 28d, and all mice died in 42d. We concluded that established model with the low EMT-6 cell concentration can provide suitable 4weeks time window for treatment , observation and test from tumor formation to dead all, and the histopathology of models’ lung was qualified with clinic features of lung cancer, clinic appearance was easy to identify , repeatability and stability test showed the models were consistent among the three times. This is more useful model for metastasis lung cancer research work.
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国内统一连续出版物号:51-1193/Q |国际标准出版物号:1000-7083
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