Application Study Of A Novel Nano NIR Probe Targeting Both EGFR And Hypoxia On Tumor Live Imaging

[Background]Tumor near infrared imaging is an essential assistant approach for tumor surgery,which need resect precisely.Traditionally,surgical locate the tumor boundary by intraoperative frozen sections,which depends on experience.Nevertheless,by this method normal tissue may be cut by mistake,causing patient’s malfunction or tumor tissue maybe omitted,causing tumor recurrence.A highly tumor-sensitive and well histocompatibility imaging molecule is required to locate the tumor boundary precisely and real-timely.However,currently,most near infrared imaging molecules targeting tumor are mono-targeting,whose sensitivity and specificity are not fine.Hence,if we can design a double-targeting near infrared imaging molecule possessing excellent sensitivity and specificity,it would be a huge beneficial for tumor surgery resection.[Objective]To design a near infrared fluorescence imaging probe based on human serum albumin,which can both target hypoxia and EGFR receptor and the probe’s ability to targeting tumor and tumor metastasis are tested through in vivo and in vitro experiment.[Methods]1.The Design and synthesis of EI-RI-CI conjugate: the connecting cite and ability between albumin and EI-RI-CI are predicted by computer molecule docking.The characteristics of HSA-EI-RI-CI and best ratio between HSA and EI-RI-CI were investigated by experiment in vitro.2.The hypoxia and EGFR-targeting of nanoprobe were investigated by western blotting and immunofluorescence experiments in vitro.3.The histocompatibility and the characteristic of nanoprobe-targeting in vivo were studied by xenograft tumor model and tumor metastasis model.[Results]1.We successfully design and synthesize a nano probe,HSA-EI-RI-CI,Which can both target hypoxia and EGFR.The stability and solubility of this nanoprobe was studied by experiment in vitro.2.The nanoprobe can significantly downregulate the expression of p-EGFR and its downstream p-AKT and p-EGFR,in HN30 and HN6,oral squamous cell carcinoma.And it can aggregate more in tumor cell in hypoxia condition than normal.3.The nanoprobe can successfully differentiate normal tissue and tumor in xenograft tumor model and tumor metastasis model.[Conclusions]1.According to the characteristic of tumor,we successfully design a nano probe,HSA-EI-RI-CI,which can both target hypoxia and EGFR receptor.The nano probe can clearly distinguish normal tissue and tumor by near infrared imaging and possess well histocompatibility.Moreover,it has excellent sensitivity and specificity of tumor-targeting,having the potential to transform to clinical application.