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尾静脉注射EMT-6细胞建立转移性肺癌模型的研究
Establishment of Metastasis Lung Cancer Model by Injection of EMT-6 Cells in BALB/c Mice
魏诗航1, 彭旭2, 韦世元1, 施雪旎1, 何学令2, 刘艳2, 尹海林2*
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作者单位:1. 四川大学生命科学学院, 生物资源与生态环境教育部重点实验室, 成都 610065;
2. 四川大学实验动物中心, 成都 610041
中文关键字:EMT-6细胞;转移性肺癌动物模型;BALB/c小鼠
英文关键字:EMT-6 cell; animal model of metastasis lung cancer; BALB/c mouse
中文摘要:目的 通过尾静脉注射EMT-6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细胞,用0.25%胰蛋白酶消化,再用磷酸盐缓冲溶液调整细胞浓度至1×106个/mL(高)、5×105个/mL(中)和1×105个/mL(低)3个浓度,尾静脉注射0.2 mL EMT-6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3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细胞注射后解剖小鼠发现,高浓度组小鼠7 d肺表面开始出现肿瘤灶,18 d内所有小鼠死亡;中浓度组小鼠7 d肺表面开始出现肿瘤灶,28 d内所有小鼠死亡;低浓度组小鼠14 d肺表面开始出现肿瘤灶,21 d所有小鼠肺表面均有肿瘤灶,荷瘤小鼠24 d开始出现死亡,35 d肿瘤灶数量达到顶峰(平均12~15个/只)且肿瘤体积变大,42 d内全部死亡。结论 相比之下,低浓度模型成瘤时间到小鼠全部死亡时间,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,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 with apparent clinical, stable and repeatable characteristics in BALB/c mice by intravenous injection of EMT-6 breast-tumor cells. Methods Cells of EMT-6 murine mammary carcinoma at logarithmic growth phase were harvested by 0.25% trypsin digestion, and equilibrated at 3 densities of1×106 cells per mL (high concentration), 5×105 cells per mL (medium concentration), and 1×105 cells per mL (low concentration) in phosphate-buffered saline. And then 0.2 mL EMT-6 tumor cells were intravenously injected. The 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 were then studied to screen the best metastasis lung cancer model. Finally, the repeatability and stability of the model were determined by 3 repeated experiments to successfully establish the metastasis lung cancer model in BALB/c mice. Results BALB/c mice were anatomized after EMT-6 tumor cells injection, and the results showed that in the group of high concentration treatment, tumor nodes on lung surface began to appear at day 7 and all animals died in 18 d; in the group of medium concentration, tumor nodes on[LM] lung surface began to appear at day 7 and all animals died in 28 d; in the group of low concentration, tumor nodes on lung surface appeared partially at day 14, all mice lung surface had tumors at day 21, the number of tumors reach the highest level (an average of 12-15/mouse), the volume of tumors became much bigger at day 35, mice began to die at day 24, and all mice died in 42 d. Conclusion We concluded that the establishment of mice models using low EMT-6 cell concentration can provide suitable 4-week time window for treatment, observation and test from tumor formation to dead, the histopathology of models' lung was consistent with clinic features of lung cancer, the clinic appearance was easy to identify, and the repeatability and stability tests showed the models were consistent among the 3 times.
2018,37(2): 191-196 收稿日期:2017-06-06
DOI:10.11984/j.issn.1000-7083.20170178
分类号:Q95-33
基金项目:四川省实验动物共享服务平台建设项目(2015JCPT0007)
作者简介:魏诗航(1993-),硕士研究生,研究方向:实验动物学,E-mail:13008199261@163.com
*通讯作者:尹海林,E-mail:13183830576@163.com
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