An Experimental Study Of MRI Targeted Diagnosis Of Hepatocellular Carcinoma By Hab18Coupled SPIO

Objective1. To explore the method and effect of the combination of human hepatoma cell-specific antibodies Hab18and superparamagnetic iron oxide (SPIO).2. Under magnetic resonance imaging (MRI) examination, to demonstrate the possibility of specific diagnosis of liver cancer by using Hab18coupled SPIO in human hepatoma cell transplantation model in nude mice.Method1. SPIO Preparation:Applying the chemical coprecipitation to synthesize the Fe3O4nanoparticles.2. Hab18-SPIO Preparation:The Hab18-SPIO was prepared through hepatocellular carcinoma (HCC) monoclonal antibodies Hab18conjugated to SPIO by using chemical crosslinking method.3. To explore the targeted effect of the Hab18-SPIO targeted contrast agent on hepatoma cell:Hepatoma HepG2cells and breast cancer cells Mcf7were adjusted to1×104/ml,1×105/ml,1×106/ml,1×107/ml, and added the Habl8-SPIO (adjusted Fe concentration to20ug/ml),after incubated1h, and then transmission electron microscope and MRI scanning.4. Construction of human human liver cancer and breast cancer xenografts in nude mice subcutaneously double, as follows:extraction0.1ml hepatoma cell fine Cell suspension (a density of1x106/ml), the anterior abdominal wall in the nude mice subcutaneously injected slowly; be taken as0.1ml, breast cancer cell suspension (a density of1x106/ml), in nude mice subcutaneously in the anterior chest wall slowly symmetrical position Bolus.20nude mice involved in molding, there are18two-tumor model in nude mice model successfully obtained36tumor mass, be divided into two groups:real group of18mass inspection by hepatoma cells were formed in nude mice subcutaneously in the anterior abdominal wall; the control group18Mass, formed by the breast cancer cell line, in the same symmetry only the anterior chest wall of nude mice subcutaneously. Via the tail vein of nude mice into the Hab18-SPIO and parallel MR scans before and after enhanced MRI measurement of the MRI signal intensity mass, calculate the rate of change of signal mass. The results of the data into statistical software16.0SPSS line statistical analysis.Strip tumor mass into slices, Prussian blue staining and HE staining.Results1. Hab18-SPIO characteristics Habl8with SPIO binding rate was93.6%measured by the Peterson method,shape regular in electron microscope,diameter of about30±5nm measured by dynamic light scattering method, Hab18successful coupling SPIO confirmed by Zeta potential detection.2. In vitro results showed that Prussian blue staining showed hepatoma cells seen in the existence of a large number of blue iron particles, breast cells significantly reduced the blue iron particles;In vitro MRI imaging showed hepatoma cells and Hab18-SPIO T2WI signal intensity were decreased after incubation, and with the number of cells increased T2WI signal level showed a decreasing trend, with statistical significance; After breast cancer cells and Hab18-SPIO incubated, the T2WI signal intensity is also reduced, and the increase in magnitude as the cells ladder by T2WI signal, but not statistically significant.3. Animal experiments Successful constructed the human human liver cancer and breast cancer xenograft in nude mice subcutaneously double model. MRI imaging in vivo showed that after injected Hab18-SPIO, the T2WI signal intensity and the change rate of the transplanted tumor has significantly decreased, with statistical significance. And calculated Hab18-SPIO for the sensitivity of tumor mass specificity Spe=77.8%, Sen=83.3%.Pathological examination:HE staining confirmed that the construction of in nude mice subcutaneously double is according to human hepatocellular carcinoma and breast cancer. Prussian blue staining showed that the large number of blue-stained iron particles in transplanted liver tumor mass organizations, only a small amount of iron particles in the breast lumps.Conclusion1. Can be constructed by chemical crosslinking method for hepatoma Hepg2cell antigen targeted MR contrast agent Hab18-SPIO.2. Prepared Hab18-SPIO targeting MR contrast agent on human hepatoma Hepg2cells have good display in vitro.3. Prepared Habl8-SPIO targeted MR contrast agents possess the necessary targeting for the nude mice transplanted liver tumor.