荧光素酶基因标记乳腺癌细胞系鉴定

目的 利用三阴性人乳腺癌MDA-MB-231细胞,对稳定表达荧光素酶的细胞系MDA-MB-231-Luc进行功能评价及鉴定.方法 构建荧光素酶的HIV慢病毒载体,体外感染乳腺癌MDA-MB-231细胞,经嘌呤霉素筛选后得到稳定表达的细胞系,采用MTT法,利用Transwell侵袭模型,应用伤口愈合细胞划痕实验评价细胞增殖、侵袭和转移能力.结果 利用慢病毒转染的MDA-MB-231细胞能稳定表达荧光素酶,与亲本细胞相比,细胞的增殖、侵袭、迁移能力无显著影响(P>0.05).结论 慢病毒感染乳腺癌细胞能稳定表达荧光素酶,对MDA-MB-231细胞的增殖、侵袭、转移能力无明显影响,为后续的小动物活体成像实验提供细胞模型.

Establishment of Luciferase-labeled Breast Cancer Cell for in vivo Bioluminescence Imaging.

Objective To establishment of triple-negative human breast cancer MDA-MB-231 with stable expression of luciferase by lentivirus vector. Methods Human breast cancer cell line MDA-MB-231 which can stably express luciferase was infected by HIV lentivirus and screened out by puromycin. The proliferation,invasion and migrasion abilities of MDA-MB-231-Luc cells were detected by MTT assay, transwell invasion assay and wound healing assay. Results Luciferase transfected into MDA-MB-231 by lentivirus vector can be stable expression. Analysis experiment results demonstrated that MDA-MB-231-Luc cell line showed similar ability of prolifetation, invasion and migration to the MDA-MB-231 cells(P>0.05). Conclusion The luciferase gene can be stably transfected into MDA-MB-231 cell by lentivirus.There is no significant effect on biological behavior with MDA-MB-231 and applied to animal bioluminescence imaging in vivo.