Construction of lung tumor cells using RFP and Fluc dual-labeled system

Abstract

Objective To establish a human lung cancer cell line that can stably express firefly luciferase (Fluc) and red fluorescent protein (RFP) gene so as to lay the foundation for the further establishment of a live-imaging lung cancer xenograft model in nude mice and therapeutic research. Methods The lentiviral vector pHBLV-Fluc-RFP containing luciferase and red fluorescent protein was constructed and then transfected into 293T cells for virus packaging. The complete virus was used to infect human lung cancer cell lines A549, H1975 and human B-cell lymphoma cell line K562. The stable cell lines were obtained by puromycin selection. Fluorescence microscopy and quantitative PCR were used to confirm the RFP and Fluc expression. Results The lentiviral vector pHBLV-Fluc-RFP was successfully constructed. Cancer cell line A549, H1975 and K562 stably expressing Fluc and RFP was obtained. The real-time quantitative PCR results showed that the relative expression of Fluc gene in the three stable infected cells was much higher than that in the corresponding wild-type cells, and the differences were statistically significant(all P<0.05). Conclusion The human lung cancer cell line A549, H1975 and human B-cell lymphoma cell line K562 with dual expression of RFP and Fluc were obtained, which provided a new model of fluorescent cells for in vivo imaging of immunodeficient mouse models such as nude mice. Key words: Red fluorescent protein; Firefly luciferase; Lentivirus; Lung cancer cell fluorescence model