抑制小鼠乳腺肿瘤转移microRNA的筛选  

The study of screening breast tumor suppressor microRNA in mice

作　　者：张丽[1] 李小颖[1] 孙彩显 张旭[1] 陈炜[1] 张连峰[1] 

机构地区：[1]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,北京100021

出　　处：《中国比较医学杂志》2014年第3期72-77,I0005,共7页Chinese Journal of Comparative Medicine

基　　金：国家"重大新药创制"科技重大专项课题"啮齿类研发平台创新药物研究开发技术平台建设"(2011ZX09307-302);国家科技支撑计划课题"神经和代谢疾病基因工程模型的建立"(2012BAI39B02)

摘　　要：目的鉴定25种microRNA的功能及其在乳腺癌中发挥的作用,以筛选新的抑制乳腺癌转移的microRNA分子。方法利用脂质体2000将25种鼠源microRNA表达载体转染至4TO7细胞,经G418筛选结合流式细胞仪分选获绿色荧光细胞得稳定表达鼠源microRNA的细胞株。将细胞2×105个/只尾静脉注射接种于BALB/C小鼠,14 d后解剖分离肺组织,统计小鼠肺组织结节数目。结果和接种阴性对照细胞小鼠相比,接种mir-449a稳转细胞的小鼠肿瘤肺转移减少。而接种mir-1935稳转细胞的小鼠肿瘤肺转移增多。其它23种microRNA稳转细胞接种小鼠肿瘤肺转移既不增加也不减少。结论从25种鼠源microRNA中筛选到2种与乳腺癌肿瘤转移相关的microRNA:mir-449a抑制乳腺癌细胞肺转移,mir-1935则促进癌细胞肺转移。Objective To investigate the functional role of 25 microRNAs in breast cancer ,and to find new tumor suppressor microRNAs that may serve as specific targets of new gene therapies . Methods Twenty-five microRNAs expression vectors were constructed and stably transfected into mouse mammary tumor cells 4To7 by Lipofectamine2000. Cells were selected with G418 and sorted by Flow cytometry.The cells in logarithmic phase were collected and 2 ＆#215;105 cells/mouse was inoculated into BALB/c mice via tail vein .Lungs were harvested 14 days after tumor cell inoculation , and the number of metastasis foci was counted .Results Mice inoculated with mir-449a-expressing 4To7 cells via tail vein developed reduced lung metastases compared with mice inoculated with negative control cells .Mice inoculated with mir-1935-expressing 4To7 cells via tail vein developed increased lung metastases compared with mice inoculated with negative control cells .Other twenty-three microRNAs neither promoted nor inhibited lung metastases of breast cancer .Conclusions Two of twenty-five microRNA were identified to be associated with breast cancer metastasis .MiR-449a may play a tumour suppressor role in the regulation of migration and metastasis in breast cancer .miR-1935 transgenic over-expression promoted tumor growth and metastasis .

关 键 词：4TO7 乳腺癌 转移 尾静脉注射 

分 类 号：R332[医药卫生—人体生理学]