Fluorescent and quantitative mitochondrial redox imaging of tumor targeted by Octa-RGD probe

摘要

Integrins,over-expressed in a broad range of cancer diseauses,are widely utilized as a tumor biomarker.Metabolism investigation also plays important roles in tumor theranostics.Devel-oping simple integrin-targetting probe and monitoring tumor metabolism will give opportunities to find ways for cancer treatment,however,the investigation of tumor metabolism with integrin receptor based probes has been rarely reported so far.Here,we developed an octavalent fuo-rescent probe Octa-R.GD by convenient genetic method,based on one tetrameric far-red fluo-rescent protein(fRFP)linked with RGD pept ides.We validated its inter gin targeting by confocal imaging in vitro.Then we screened a variety of tumor cells,and differentiated their binding affinity based on the fuorescence of the probe via flow cytometry.Among these cells,CNE-2 cells had the highest uptake of the probe,while B16 cells had the lowest,corresponding with their intergin expression levels.Next,the fuorescent and metabolic imaging was performed in HT1080(intergin postive)tumor,where nicotinamide adenine dinudeotide hydrogen(NADH),flavo-protein(Fp)and fRFP fuorescent signals were collected.The tumor from mice intravenously injected with Octa RGD probe displayed obviously higher NADH redox ratio NADH/(Fp+NADH)and fRFP signal,than those with fRFP protein.It suggested that integrin targeting may have influence on the target cell metabolism,and further demonstrated Octa-R.GD probe facilitated its uptake in the targeted tumor in vrivo.This paper developed a useful probe,which can bind integrins speci-cally and e±ciently in tumor cells,and together with tumor metabolic information,it may provide new insight for RGD targeting-based cancer therapeutics.