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李小颖,马元武,张旭,张连峰.不同接种量荧光素酶标记小鼠乳腺癌细胞4T1在小鼠体内生长及肺转移的比较[J].中国实验动物学报,2012,(1):14~17.
不同接种量荧光素酶标记小鼠乳腺癌细胞4T1在小鼠体内生长及肺转移的比较
Comparison of the tumor growth and pulmonary metastasis in mice transplanted with different number of luciferase-labeled mouse breast cancer 4T1 cells
  
DOI:10.3969/j.issn.1005-4847.2012.01.004
中文关键词:  4T1小鼠乳腺癌细胞株  荧光素酶  肺转移  肿瘤模型
英文关键词:Mouse mammary tumor 4T1 cell line  Luciferase  Pulmonary metastasis  Tumor model
基金项目:卫生部项目,实验动物和人类疾病动物模型资源扩展(200802036)和十一五新药专项支持(2008ZX09305-001)。
作者单位E-mail
李小颖 中国医学科学院,北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021 lixiaoyinghebei@163.com 
马元武 中国医学科学院,北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021  
张旭 中国医学科学院,北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021  
张连峰 中国医学科学院,北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021  
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中文摘要:
      目的采用活体成像技术比较四种剂量荧光素酶标记肿瘤细胞在小鼠体内生长及肺转移情况,为光学标记肿瘤模型的药物筛选或机制研究提供参考资料。方法以荧光素酶作为报告基因导入小鼠乳腺癌细胞4T1中,经G418筛选获得稳定表达荧光素酶的细胞克隆并扩大培养。标记细胞稀释成1×107 细胞/mL, 2×107细胞/mL, 5×107细胞/mL和1×108细胞/mL四种剂量,取0.1 mL接种于BALB/c小鼠右侧第二对乳腺脂肪垫内,制作小鼠原位乳腺癌模型,比较肿瘤细胞在小鼠体内生长及肺转移情况。结果获得稳定表达荧光素酶基因的细胞克隆,在致瘤性方面和亲代细胞无明显差别,四种剂量细胞接种BALB/c小鼠后,均有肿瘤生长,接种第28天时,四种剂量接种的原位移植瘤大小没有明显差别,但接种两个高剂量肿瘤细胞的小鼠组各有2只小鼠死亡;接种后31 d,发现四种剂量接种的原位移植瘤均发生不同程度的转移,随着观察天数的增加,转移程度逐渐严重,接种后42 d,小鼠陆续发生死亡。结论根据转移和死亡情况,确定接种1×106个细胞/只不仅肺转移明显,而且存活时间一般超过45 d,比高剂量接种存活时间长,为最佳肺转移剂量。
英文摘要:
      ObjectiveTo study the tumor growth and pulmonary metastasis in mice transplanted with different number of luciferase-labeled mouse breast cancer cells, and to provide important information for drug screening and cancer mechanism research. MethodsThe vector containing luciferase gene was constructed and transfected into mouse breast tumor cell line 4T1 cells and selected with G418 to obtain stable Luc-expressing clones. The cells in logarithmic phase was collected and diluted to 1×108,5×107,2×107 and 1×107 cells/mL. Then 0.1 mL cell suspension was inoculated into the second mammary fat pad on the right side of normal BALB/c mice to establish the tumor models. Their tumorigenesis and metastasis were analyzed in vivo.ResultsThe stable Luc-expressing cell lines were obtained which had no obvious difference with parental cells in tumorigenicity. The whole-body optical imaging found that tumor could be formed after the cells were inoculated orthotopically. After 28 days, the tumor sizes were similar among the four concentrations, while there were two mice each died at 5×106 and 1×107 cells concentrations. After 31 days, the tumor metastasis showed different degree among the four concentrations. The metastasis became more seriously with time passed. However, after 42 days, deaths of mice occurred in succession.ConclusionsThe number of 1×106 cells is the best concentration, not only induces obvious pulmonary metastasis but also shows a survival time longer than 45 days, compared with that induced at other higher concentrations. 
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