稳定表达荧光素酶的人乳腺癌MCF-7细胞的构建

Establishment of the human breast cancer MCF-7 cells line stably expressing luciferase

目的构建稳定表达荧光素酶的人乳腺癌MCF-7细胞株。方法采用脂质体转染法将荧光素酶基因导入MCF-7细胞,经潮霉素B筛选建立稳定表达荧光素酶的MCF-7-Luc细胞株;将不同数量MCF-7-Luc细胞分别接种于黑色96孔板(细胞数量59~30000)和裸鼠皮下(细胞数量2.44×10^(3)~5×10^(6)),IVIS LuminaⅢ成像系统体外培养细胞以及裸鼠体内荧光素酶表达效率。结果筛选得到稳定表达荧光素酶的MCF-7-Luc细胞;IVIS LuminaⅢ成像系统检测结果表明,不论是体外培养的细胞或是裸鼠体内,生物发光强度与接种的细胞数量之间均存在明显的线性关系(体外y=129.32x-26409,R^(2)=0.9922,P<0.05;体内y=106.82x+1E+07,R^(2)=0.9404,P<0.05)。结论成功构建了稳定表达荧光素酶的MCF-7-Luc细胞,为研究乳腺癌的发生发展、肿瘤微环境以及抗乳腺癌药物的筛选提供必要条件。

Objective To establish MCF-7 cell strains stably transfected with Luciferase gene(Luc).Methods MCF-7 cells were transfected by lipidosome containing luciferase gene and screened by hygromycin B for establishing MCF-7-Luc lines that could stably express the luciferase.MCF-7-Luc cells were inoculated into the 96-well plate(59~30,000)or inoculated subcutaneously to nude mice(2.44×10^(3)~5×10^(6)),and the expression of reporter gene of luciferase was detected by IVIS LuminaⅢBiological Live Imaging System.Results MCF-7-Luc cells that stably expressed luciferase were screened out.Bioluminescence intensity was correlated in a linear manner with the number of MCF-7-Luc cells in vitro or in vivo(for in vitro correlation,y=129.32x-26409,R^(2)=0.9922,P<0.05;for in vivo correlation,y=106.82x+1 E+07,R^(2)=0.9404,P<0.05).Conclusion MCF-7 cell lines with stable expression of luciferase were established.It will provide conditions for the study of the occurrence and development of breast cancer,and the development anti-breast cancer drugs.