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.