A Dual-Targeting Functionalized Graphene Film for Rapid and Highly Sensitive Fluorescence Imaging Detection of Hepatocellular Carcinoma Circulating Tumor Cells.

High recurrence and metastasis rates are the major causes of the high mortality of hepatocellular carcinoma (HCC). Circulating tumor cells (CTCs) disseminating into the bloodstream plays an essential role in cancer metastasis. However, since HCC-CTCs are extremely rare, limitations of current detection methods impede accurate discerning HCC-CTCs under complicate biological context. Here, a dual-targeting functionalized rGO film (DTFGF) for specific identifying HCC-CTCs was created via coinstantaneous targeting epithelial cell adhesion molecule (EpCAM) and HCC cell-specific asialoglycoprotein receptor (ASGPR). Anti-EpCAM antibodies and galactose-rhodamine-polyacrylamide nanoparticles (Gal-Rh-PAA NPs) specifically recognizing ASGPR are modified on the surface of a graphene film that quench the rhodamine fluorescence. HCC-CTCs can be captured by anti-EpCAM antibody and endocytose Gal-Rh-PAA NPs, recovering the rhodamine fluorescence. Profiting from accuracy of dual-targeting, less handling steps and high resolution of fluorescence imaging, a simple, rapid and low-cost HCC-CTC enumeration method is established with excellent sensitivity and selectivity than conventional methods. Using DTFGFs, as low as 5 HCC-CTCs were detected in a 1 mL blood sample. Further results revealed that larger HCC-CTCs quantities indicate more advanced stages of HCC in patients. Overall, this work holds great promise for the early diagnosis, prognosis and therapeutic evaluation of HCC.