Competitive electrochemical platform for ultrasensitive cytosensing of liver cancer cells by using nanotetrahedra structure with rolling circle amplification

摘要

In this work, a competitive and label-free electrochemical platform was performed for the ultrasensitive cytosensing of liver cancer cells based on DNA nanotetrahedron (NTH) structure and rolling circle amplification (RCA) directed DNAzyme strategy. The multifunctional nanoprobes were fabricated through a DNA primer probe, carboxyfluorescein (FAM) functionalized TLS11a aptamer and horseradish peroxidase (HRP) immobilized on the surfaces of the platinum nanoparticles (PtNPs). Then the NTH-based complementary DNA (cDNA) probe, complementary to the TLS11a aptamer, was attached on a disposable screen-printed gold electrode (SPGE) for increasing the reactivity and accessibility with the prepared nanoprobes. Due to the primer probe and the circular probe with G-quadruplex sequences for RCA, it can lead to the formation of numerous G-quadruplex/hemin DNAzyme, thus generating a remarkable electrochemical response. When the target cells were present, the nanoprobes were released from the SPGE due to the specific recognition of TLS11a aptamers for HepG2 cells, resulting in the electrochemical signal changes. The cytosensor was ultrasensitive for HepG2 tumor cell detection with a detection limit of 3 cell per mL. Furthermore, this strategy was also demonstrated to be applicable for cancer cell imaging. In summary, this electrochemical cytosensor holds great potential for circulating tumor cell detection in the early cancer diagnose.