Zell- und Molekularbiologie

Abstrakt

Luciferase Expression is more Accurate than GFP to Assess Mirnas- Relevant Oncogenesis in vivo Live Imaging Study

Background: In vivo live imaging technology was able to assess dynamically tumor processes and biology throughout the entire experiment period depending on a bioluminescent signal from tumor cells generated from expression of Green Fluorescent Protein (GFP) or the firefly luciferase gene. Recently, there is scarely reported a comparison of Luc- and GFP-transfected tumors to assess the biological mechanism of miRNAs using identical model systems.

Methods: We constructed the SGC-7901 cells which involving stability-enhanced Luc- and GFP-coexpression identified by real time PCR (qPCR) and fluorescence microscope, the constructed cells were injected into the nude mice, then we compared the transfection efficiency and accuracy of GFP and luciferase for miRNAs-relevant oncogenesis after 40 days.

Results: We constructed successfully SGC-7901 cell lines coexpressing the stability-enhanced luciferase and GFP. The miR-145 expression of Luc/LV-miR-145 SGC-7901 cells was higher than Luc/LV-ctrl’s, and luciferase intensity was consistent with tumor volume and negatively related to miR-145 expression in Luc-transfected SGC-7901 cells, but the phenomenon of GFP was opposite. Conclusion: We believed that luciferase offers distinct advantages over GFP as a transfection and gene expression reporter to assist the studies of miRNAs-relevant oncogenesis in vivo live imaging studies.