一种易于循环肿瘤细胞研究的4T1-G乳腺癌稳定细胞株的构建  

Construction of 4T1-G stable breast cancer cell line for the study of circulating tumor cells

作　　者：刘淑荣 丁怡[1] 万晓庆[1] 成丛丛 王艺斐[1,2] 史立宏[1,3] 路中[1,2] 王丽华[1,2] 王学健[1,3] LIU Shurong;DING Yi;WAN Xiaoqing;CHENG Congcong;WANG Yifei;SHI Lihong;LU Zhong;WANG Lihua;WANG Xuejian(Department of Molecular Oncology Laboratory,Weifang Medical University,Weifang 261053,China;the Affiliated Hospital of Weifang Medical Untverstty;Department of Pharmacology Laboratory,Weifang Medical University)

机构地区：[1]潍坊医学院分子肿瘤学实验室,山东潍坊261053 [2]潍坊医学院附属医院 [3]潍坊医学院药理学教研室

出　　处：《潍坊医学院学报》2018年第3期184-187,共4页Acta Academiae Medicinae Weifang

基　　金：国家自然科学基金资助课题(课题编号:81472489);山东省科技发展计划项目资助课题(课题编号:2014GGB14403)

摘　　要：目的构建一种易于研究乳腺癌循环肿瘤细胞(CTCs)过表达绿色荧光蛋白GFP的4T1细胞株。方法通过慢病毒感染,使小鼠三阴性恶性乳腺癌4T1细胞稳定过表达GFP,构建4T1细胞株(4T1-G);将该细胞接种于BALB/c小鼠乳腺脂肪垫内,制作小鼠4T1-G乳腺癌原位移植荷瘤模型,应用免疫磁珠分选结合GFP荧光方法检测动物模型外周血中的循环4T1-G细胞。结果经慢病毒感染过表达GFP的4T1-G细胞其迁移与侵袭能力及上皮间质转化(EMT)表型标志物的表达与亲本4T1细胞无显著性差异。4T1-G细胞接种BALB/c小鼠后,在接种部位观察到肿瘤生长,通过免疫磁珠分选结合GFP荧光成功分选小鼠乳腺癌模型外周血中的循环4T1-G细胞。结论 4T1-G乳腺癌GFP稳定表达细胞株不仅其荧光特性易于CTCs的确认,而且保持亲本细胞的EMT表型和侵袭特性,因此,4T1-G乳腺癌GFP稳定表达细胞株的构建为开展CTCs及肿瘤转移的整体动物研究提供一个良好的模型。Objective To construct a green fluorescent protein（GFP） positive breast cancer stable cell line for the study of circulating tumor cells（CTCs）. Methods By the lentivirus infection,a GFP-positive 4 T1 mouse triple negative malignant breast cancer stable cell line（4 T1-G） was constructed. It was established the mouse 4 T1-G breast cancer orthotopic model by inoculating cells in BALB/c mouse mammary fat pad. The circulating 4 T1-G cells in peripheral blood were detected by immunomagnetic beads separation combined with GFP fluorescence. Results There was no significant difference in the expression of phenotypic markers between 4 T1-G cells infected with lentivirus and parental 4 T1 cells in their migration and invasion ability and epithelial interstitial transformation. After the cells were inoculated with BALB/c mice,the growth of tumors were observed at the inoculated site,and circulating 4 T1-G cells in peripheral blood in mice were successful detected by immunomagnetic bead cell sorting combined with GFP fluorescence. Conclusion 4 T1-G stable breast cancer cell line not only has the green fluorescence facilitative the confirmation of CTCs,but also keeps the original EMT phenotypes and invasiveness characteristics of the parental cells. 4 T1-G cell line is a good model for investigating CTC and metastasis in vivo.

关 键 词：乳腺肿瘤 绿色荧光蛋白 循环肿瘤细胞 肿瘤转移 

分 类 号：R34[医药卫生—基础医学]