The Study Of CRGD Nanoparticles With Blocking Tumor Vessel Function On Ovarian Cancer

ObjectiveTo develop anti-ovarian cancer cell proliferation and invasion ability,and to block the multi-target nanoparticles of ovarian cancer blood supply.To provide more valuable experimental evidence for the clinical treatment of ovarian cancer.Methods1.The cytotoxic effects of low molecular weight heparin and folic acid on SKOV3 and OVCAR-3 of ovarian cancer cells were investigated by MTT method.2.The chemical modification of low molecular weight heparin for carboxyl heparin,and folic acid in accordance with different mole ratio of chemical conjugation to carboxyl on heparin,the preparation of 4 kinds of different proportion of heparin-folic coupling,by determined by MTT method,Transwell method,three-dimensional cultivation experiment investigation on ovarian cancer cell SKOV3 at a cellular level and OVCAR-3 targeting ability,inhibition of proliferation,invasion and microtubule formation ability,so as to screen out the best proportion of heparin and folic acid solution,get the best effect of heparin-folic normalize anticancer drug delivery system model.3.In the previous step of heparin-folic acid in the drug delivery system,further chemical access different molar ratio of RGD peptides can block tumor blood supply,preparation with nanoscale coupling RGD peptides of heparin-folic-RGD drugs.By determined by MTT method,Transwell method,three-dimensional cultivation experiment investigation on ovarian cancer cell SKOV3 at a cellular level and OVCAR-3 targeting ability,proliferation and inhibition of microtubule formation ability,compare two kinds of heparin-folic-RGD nano drug effects.The optimum RGD nanoparticles were selected for further vivo experiments.4.To establish animal model of ovarian cancer through in vivo imaging of the heparin-folic-RGD nano drug distribution in vivo biological,and through the monitoring of tumor size,immunohistochemical CD34,Ki67,CD34-PAS methods such as the in vivo tumor suppression effect was observed.Results1.Heparin can have cytotoxic effects on SKOV3 and OVCAR-3 of ovarian cancer cells in specific drug concentration,but it has certain drug concentration threshold.When the drug concentration continued to improve,the effect was not obvious.And as a natural macromolecular heparin,has good biocompatibility and degradability,especially in tumor tissues with the characteristics of permeability and retention(EPR),so as a carrier of the drug,heparin can be combined with anticancer drugs,design new drug system.2.Ovarian cancer cells SKOV3 and OVCAR-3 were both high expression cell lines of folic acid receptors;The toxic effect of folic acid on these two cells is dependent on concentration,that is,with the increase of the concentration of folic acid,the more obvious the cytotoxicity,the better the inhibition effect on the proliferation of ovarian cancer cells.So folic acid can be used as a targeted drug and treatment for ovarian cancer.3.The different molar ratio of folic acid and heparinization coupling form the duality of drug delivery system,under the same quality,with the increase of the proportion of folic acid,the ovarian cancer cell proliferation,invasion,and inhibition of microtubule formation ability enhancement.Therefore,HF20 can be screened to optimize the drug delivery system.4.The HF20 as nanodrugs skeleton,further chemical coupling can inhibit effect on ovarian cancer cell proliferation and block the ovarian blood supply of RGD peptides,design and preparation of nano-sized drug heparin-folic-RGD,by cellular level experiments,screening treatment work best HF20R4 for the final system of nanodrugs.5.In the animal body level,it was further verified that the HF20R4 nano-drug system had a good drug targeting and retention effect(EPR)on SKOV3 tumor-bearing nude mice through drug distribution experiments.By examining each animal body weight,volume of subcutaneous tumor growth,tumor tissue of immunohistochemical experiment,verified the nano drug to inhibit ovarian cancer cell proliferation and tumor endothelial dependent blood vessels(EDV)and vascular mimicry(VM)blood supply,so as to achieve the effect of the treatment of ovarian cancer.Conclusion:This study more targets of the preparation of nano drug,can restrain the proliferation of ovarian cancer cells,can block the tumor blood supply at the same time,to guide the clinical treatment of ovarian cancer provides more valuable experimental basis.