Folic acid modified copper nanoclusters for fluorescent imaging of cancer cells with over-expressed folate receptor.

Abstract

Water-soluble and functional copper nanoclusters (CuNCs) were prepared by using folic acid (FA) that serves both as a reducing reagent and a stabilizer. FA also acts as a functional ligand on the surface of the CuNCs, and this can be exploited to target the folate receptor which is over-expressed on the surface of HeLa cells. The FA-modified CuNCs nanoclusters have an average size of ca. 0.9nm and are stable in aqueous medium for 30days. Under photoexcitation at λex 270 and 350nm, the FA-CuNCs display strong blue fluorescence with an emission peak at 440nm. The FA-CuNCs exhibit low cytotoxicity and favorable biocompatibility as demonstrated by an MTT assay. A cell viability of >80% is found when incubating HeLa cells for 20h with FA-CuNCs at levels of up to 200μgmL-1. The targeting capability of the FA-CuNCs is demonstrated by live cell imaging. It is shown that HeLa cells with over-expressed folate receptor are much brighter than A549 cells where the receptor is not over-expressed. This is further corroborated by the fact that the copper content in HeLa cells (1.5 pg/cell) is 6.5-fold higher than that of A549 cells (0.23pg/cell), both measured after the same incubation time of 3h. If free FA is introduced into the cell culture medium, the folate receptors will be preoccupied with FA, and this results in a significant decrease in the cellular uptake of the FA-CuNCs by HeLa cells. Graphical Abstract Biocompatible copper nanoclusters (CuNCs) coated with folic acid (FA) were prepared and are shown to be viable probes for the differentiationbetween FR-positive HeLa cells and FR-negative A549 cells.