Synthesis Of Targeted Drug-loaded Nanoparticles And A Preliminary Study On Their Anticancer Effects To Human Pancreatic Cancer Cell

Carcinoma of the pancreas is one of the leading causes of death from malignant disease with poor prognosis. Although enormous efforts have been made in early detection and comprehensive treatment for this disease,there has been only marginal improvement in the outcome of treatment for pancreatic cancer. Chemotherapy received widespread attention as one of the treatment for pancreatic cancer. But the poor bioavailability and toxicity are the key issues affecting the anticancer treatment.Nanotechnology can play a pivotal role by delivering drugs in a targeted fashion to the malignant cells that will reduce the systemic toxicity of the anticancer drug. In this report, we selected epidermal growth factor receptor(EGFR) as a molecular target for pharmacologic intervention and conjugated EGFR specific peptide to the particle surface of nanoparticles in order to actively target pancreatic cancer cell line. As the result, we show a stepwise development of a nanoparticle-based targeted delivery system for therapeutic application in pancreatic cancer.We obtained( 25±7.6),(49±8.6),(88±20.5),(145±13.5),(205±8.4) nm ICG-PBCA-NPs by regulating the temperature, the stirring speed,the amount of stabilizer and raw materials during the emulsion polymerization. The absorbency(A value) of ICG solution was measured at 805 nm with ultraviolet/visible specrtromete. The standard curve of ICG was drawn according to the relationship between the concentration and the A value. Mice were randomly divided into 7 groups.Livers, kidneys, spleens, hearts and pancreas removed from mice weresacrificed for frozen section fluorescence examinations at 1, 5 and 12 hours.At the same time, The frozen section’s fluorescence values were examined when obtaining the samples from organs at 0.5, 1, 5 and 12 hours after the injection respectively. As a result, the group(25±7.6) nm ICG-PBCA-NPs showed no obvious distribution in each tissue of mice. The pancreatic tissues’ has the strongest fluorescence intensity in group(49 ± 8.6) nm.While livers and spleens’ strongest fluorescence intensities appeared in(145±13.5) nm size. Fluorescence intensity changed little in heart with the increase diameter of nanoparticle. The strongest fluorescence of various tissues shortened as the nanoparticles size decreases.Epidermal growth factor receptor targeted polybutylcyanoacrylate nanoparticle carrying 5-Fu(5-Fu-EGFR-NPs) are developed by conjugating a single-chain anti-EGFR antibody to the surface of functionalized polybutylcyanoacrylate nanoparticles with the method of emulsion polymerization. Firstly we studied the immunal activity of5-Fu-EGFR-NPs by enzyme-linked immunosorbent assay. Secondly MTT assay was performed to estimate the effects of 50, 40,30,20, 10, 5 ug?m L-1unloaded polybutylcyanoacrylate nanoparticle(PBCA-NPs), 5-Fu loaded polybutylcyanoacrylate nanoparticles(5-Fu-PBCA-NPs) on pancreatic cancer cells at 12, 24, 48 h. Thirdly we used the formula to calculate the cell survival rate and inhibition rate. As a result, PBCA-NPs as targeted therapy vectors have biosafety.The inhibition rate of 5-Fu-PBCA-NPs and5-Fu-EGFR-NPs were different significantly at 24, 48h(P<0.05, P<0.01).The inhibitory effect of bare drug 5-Fu group stronger than5-Fu-PBCA-NPs group at 12, 24h(P<0.05, P<0.01). However, the anti-tumor effect of 5-Fu drug group weaker than the 5-Fu-PBCA-NPs group at 48h(P<0.01). In addition, the tumor suppression rate of5-Fu-PBCA-NPs, 5-Fu-EGFR-NPs and 5-Fu showed a dose-dependent.The various ranges of diameter could affect the biodistribution of nanoparticles dramatically. ICG-PBCA-NPs with a 49 nm diameter has thebest pancreas targeting. On this basis we prepared the same size5-Fu-PBCA-NPs and 5-Fu-EGFR-NPs. In vitro experiments the group5-Fu-EGFR-NPs’ cell inhibition rates were stronger than the group5-Fu-PBCA-NPs. In summary, our study indicated that EGFR targeted polybutylcyanoacrylate nanoparticles as a carrier of 5-Fu could achieve perfect anticancer effects.