title: Establishment of a Human Breast Cancer Model by Fusion PCR for In Vivo and In Vitro Fluorescence Imaging of Human Breast Cancer.
abstract: This study aimed to construct a breast cancer model that could continuously express the genes of luciferase and green fluorescent protein. The genes luciferase, EGFP, and Neo were obtained by fusion polymerase chain reaction (PCR) and inserted into pAAV-MCS. The pAAV-Luciferase-EGFP-Neo vector was transfected into MDA-MB-231 cells. After antibiotic resistance gene screening and limiting dilution assay, we constructed a monoclonal stable cell line that expresses the fusion protein Luciferase-EGFP. In comparison with the polyclonal stable cell line, the monoclonal cell line had good genetic stability and was not different from the parental cell line MDA-MB-231. The monoclonal stable cell line would be ideal for a breast cancer model. Indices of fluorescence imaging can be applied to fluorescence imaging in vitro and in vivo, providing a straightforward and reliable system for breast cancer and drug discovery research.
authors.affiliation: 1 College of Life Science, Shanghai University , Shanghai, China .|2 Suzhou Institute of Biomedical Engineering and Technology , Chinese Academy of Sciences, Suzhou, China .
authors.forename: Can
authors.initials: C
authors.lastname: Huang
authors.affiliation: 3 College of Life Science and Technology, China Pharmaceutical University , Nanjing, China .
authors.forename: Wenjun
authors.initials: W
authors.lastname: Lan
authors.affiliation: 1 College of Life Science, Shanghai University , Shanghai, China .|2 Suzhou Institute of Biomedical Engineering and Technology , Chinese Academy of Sciences, Suzhou, China .
authors.forename: Feifei
authors.initials: F
authors.lastname: Wang
authors.affiliation: 2 Suzhou Institute of Biomedical Engineering and Technology , Chinese Academy of Sciences, Suzhou, China .
authors.forename: Chun
authors.initials: C
authors.lastname: Zhang
authors.affiliation: 2 Suzhou Institute of Biomedical Engineering and Technology , Chinese Academy of Sciences, Suzhou, China .
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Liu
authors.affiliation: 1 College of Life Science, Shanghai University , Shanghai, China .
authors.forename: Qin
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000235:genetics; D051379:Mice; D061848:Optical Imaging; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008156:Luciferases
keywords: EGFP; fluorescence imaging; fusion protein; luciferase; monoclonal cell line