Study On Targeted Labeling And Hyperthermia Of Hepatoma Cells By Au@SiO₂@CuInS₂-ZnS Quantum Dots Probe

Objective:To synthesize a novel gold nanostar-silica-sulfur indium copper sulfide anti-alpha-fetoprotein antibody?Au@SiO₂@CuInS₂/ZnS-anti-AFP?quantum dot probe and explore its targeted markers and fluorescent imaging capabilities,cytotoxicity,photothermal efficiency,and hyperthermia efficacy of liver cancer cells,in order to provide a new method for the diagnosis and treatment of liver cancer.Methods:The expression of alpha-fetoprotein in hepatocellular carcinoma cell line HepG2 was explored by immunohistochemistry.Hydrothermal synthesis of Au nanostar-silica-sulfur indium copper sulfide anti-alpha-fetoprotein antibody probe and Au nano-cage sulphur indium copper sulphide anti-alpha-fetoprotein antibody probe,and compare the physical and biological performance.Laser confocal microscope was used to detect the target labeling and fluorescence imaging ability of the probe after the incubation with the Hep-G2 cell line.The MTT method was used to detect the cytostatic rate to evaluate the biocompatibility of the probe.Hep-G2 cells were incubated with probes,and the cell culture medium was irradiated with a laser of808 nm and 1.5 W/cm²,and the temperature of the cell culture solution during photothermal hyperthermia was measured with a thermometer;the apoptosis rate of the hyperthermia cells was detected by flow cytometry.Results:The results of immunohistochemistry showed that the cytoplasm and cell membrane of Hep-G2 cells were brown,the nucleus was blue,indicated that the cytoplasm and membrane strongly expressed AFP.Targeted labeling experiments:The laser confocal microscopy showed that the HepG2 cytoplasm emitted yellow fluorescence under the excitation of 520 nm wavelength light,and the nucleus emitted blue fluorescence under the excitation of 360 nm wavelength light.MTT assay showed that compared with 20%gold nanocage probe,the HepG2 cell inhibition rate of 20%gold nanostar probe were reduced significantly at 20h,12h and 24h,respectively?0.61±0.0105 VS 0.167±0.0117;0.201±0.0111 VS 0.467±0.0272;0.387±0.0446 VS 0.774±0.0375;P<0.05?.Photothermal experiments showed that the plateau temperature and the temperature rise of the 20%gold nanostar probe were significantly higher than the 20%gold nanocage probe?45.5°C,43.3°C?,the difference was statistically significant?Z=-5.895,P<0.001?.The results of hyperthermia experiments showed that the apoptotic rate of gold nanostar probe hyperthermia reached 95%-98%plateau at 40 min;it takes 50 minutes to stabilized at86%when it comes to gold nanocage probes.There was a statistically significant difference in apoptotic rate between the probes.The hyperthermia effect of gold nanostar probe was significantly higher than that of gold nanocage probe?Z=-4.846,P<0.001?.Conclusion:?1?The AFP expressed on the cytoplasm and membrane of HepG2 cell line.?2?The novel gold nanostar probe specifically targets and marks HepG2 cells by antigen-antibody reaction,and can fluorescently image the cells by fluorescence excited by quantum dots.?3?The 20%gold nanostar probe has a significantly reduced cytotoxicity compared to the 20%gold nanocage probe and with fine biocompatibility.?4?The photothermal temperature,the cell killing rate and the photothermal efficiency of 20%the novel gold nanostar probes were higher than the 20%gold nanocage probes?45.5°C,43.3°C,Z=-5.895,P<0.001??Z=-4.846,P<0.001?.