一种基于EdU的体外癌细胞与小鼠体内组织细胞标记方法的研究

An EdU-based Method for in vitro Cancer Cells and Mouse in vivo Tissues Cells Labeling

目的:比较EdU标记对三种癌细胞和小鼠对细胞增殖的影响,为EdU作为标记开展相关细胞增殖实验和临床研究提供依据。方法:本研究使用不同剂量EdU对人非小细胞肺癌A549细胞、人宫颈癌Hela细胞、人肝癌Huh7进行标记2 h,然后使用荧光显微镜观测EdU在细胞中的标记效率,并使用多波长荧光酶标仪检测这三种癌细胞系标记后的荧光强度;使用流式细胞仪检测小鼠经不同剂量的EdU干预12 h后,体内肺、肝、肾组织标记的荧光强度。结果:与对照组相比,经EdU处理后,A549和Hela细胞系的荧光强度,三个剂量组均有显著性差异(P<0.01),Huh7细胞系的荧光强度,50μmol/L有显著性差异(P<0.05);EdU在小鼠体内组织肺、肝、肾组织中均有分布,且在肝组织中分布比肺组织和肾组织高。结论:EdU的体外癌细胞与小鼠体内组织细胞的标记效率各不相同,建立的EdU体外标记癌细胞和小鼠体内组织的方法简单,易操作。

Objective:To investiagte the effects of EdU on cell proliferation after in vivo tissue labeling of three types of cancer cells and mice,and to provide basis for EdU as a marker for relevant cell proliferation experiments and clinical studies.Methods:In this study,human non-small cell lung cancer A549 cells,human cervical cancer Hela cells and human liver cancer Huh7 cells were labeled with different doses of EdU for 2 h.The labeling efficiency of EdU in the cells was observed by using fluorescence microscopy,and the fluorescence intensity of the three cancer cell lines were measured by using a multi-wavelength fluorescence enzyme standard.The fluorescence intensity of lung,liver and kidney tissue markers in vivo were measured by using flow cytometry in mice after 12 h of intervention with different doses of EdU.Results:Compared with that in the control group,the fluorescence intensities of A549 and Hela cell lines after EdU treatment were significantly different in all three dose groups(P<0.01),and the fluorescence intensity of Huh7 cell line was significantly different at 50μmol/L(P<0.05);EdU distributed in lung,liver and kidney tissues in mice in vivo,and the distribution was higher in liver tissues than in lung and kidney.The distribution of EdU was higher in liver tissues than that in lung and kidney tissues.Conclusions:The labeling efficiency of EdU varies between cancer cells and mouse in vivo tissue cells.The labeling efficiency of EdU in cancer cells in vitro and mouse in vivo tissues are different The established method for in vitro labeling of cancer cells and mouse in vivo tissues with EdU is simple and easy to operate.