Preparation Of Hollow Mesoporous Silica-Curcumin Nanoparticles And Its Effect On Proliferation,Apoptosis And Autophasy Of Hepatoma Cancer Cells

Background and objective: Curcumin is an effectively active ingredient extracted from the roots of curcuma,which is a natural and antitumor drug with wide application prospects.But the property of curcumin,including poor water-soluble and low bioavailability greatly limits the clinical promotion of curcumin.First of all,hollow mesoporous silica(HMS)nanoparticles was prepared,Which curcumin was loaded on to obtain hollow mesoporous silica-curcumin(HMS-CUR)nanoparticles.Then a series of experimental methods were used to evaluate the properties of this hollow mesoporous silica-curcumin nanoparticles.The main investigations are as follows: 1.explore the effects of hollow mesoporous silica-curcumin nanocparticles on the proliferation and apoptosis of hepatocellular carcinoma cells.2.effect of hollow mesoporous silica-curcumin nanocparticles on autophasy.This can not only solve the problem of low bioavailability of curcumin,but also can clarify the mechanism of hollow mesoporous silica-curcumin nanocparticles against hepatocellular carcinoma,which provides a theoretical basis for the clinical application of liver cancer.Methods: 1.Cationic polystyrene template(CPS)was prepared by emulsion polymerization.Silica nanoparticles was developed by the addition of Na2 Si O3 · 9H2 O with CPS as templates.HMS was made by calcinating at high temperature.Scanning electron microscopy(SEM)was used to characterize CPS and HMS.2.HMS and curcumin were mixed in a certain proportion.After mechanical stirring,HMS-CUR was prepared.The HMS-CUR was characterized by transmission electron microscopy(TEM).Fourier transform infrared spectroscopy(FT-IR)was used as a method of quality evaluation.UV-Vis spectrophotometer was used for release study in vitro.3.Four groups were put up in this experiment,which was involved in the blank control,the group of HMS,the group of free curcumin(concentration: 5μM)and the group of HMS-CUR(actual curcumin content: 3.5μM).The inhibitory effect of HMS-CUR on hepatoma cancer cells was determined by CCK-8 assays and clonogenic assays.Western Blot was used to detect the expression of proliferating protein in hepatocarcinoma cells.4.Hoechst 33342 staining was used to observe the occurrence of apoptosis after drug treatmen.Annexin V ? FITC/PI staining was performed for apoptotic rate.The apoptosis related proteins were detected by Western Blot,which is used as a semi-quantitative method of detection.5.The level of autophagy-related protein is dependent on the mesurement of Western Blot.Results: 1.The results of SEM showed that CPS and HMS were uniform in size and the particle size was between 50 and 100 nm.2.Both of TEM and FT-IR confirmed that curcumin was loaded on HMS.UV-Vis analysis revealed the rapid release of HMS-CUR in ethanol and slow release in DMEM,10% FBS-DMEM.3.Both of CCK-8 assays and clonogenic assays indicated that HMS-CUR had a significant inhibitory effect on hepatoma cancer cells(p<0.05).Moreover,HMS-CUR revealed a sustained-release effect.HMS-CUR can significantly down-regulated the expression of proliferating cell nuclear antigen(PCNA)(p<0.05).4.The results of Hoechst33342 assay and Flow cytometry demonstrated that HMS-CUR was able to induced apoptosis in hepatoma cancer cells(p<0.05).Western Blot manifested that HMS-CUR can obviously up-regulated the expression of apoptosis related proteins(Cleaved-PARP,Cleaved-Caspase3)(p<0.05).5.HMS-CUR also activated autophasy,which can be evidenced by a higher ratio of LC3-I to LC3-II and up-regulated expression level of Beclin1(p<0.05).Additionally,HMS-CUR notbly suppressed the level of P62 / SQSTM1 in SMMC-7721 cells(p <0.05),but potentiated the expression of P62 / SQSTM1 in Hep G2 cells conversely(p <0.05).Conclusions: As a new drug delivery system with characteristics of non-toxic and good biocompatibility,HMS effectively solves the problem of poor water solubility of curcumin.Compared with free curcumin,HMS-CUR can notably inhibit the proliferation of hepatocarcinoma cells,induce apoptosis and activate autophagy.Remarkably,HMS-CUR has a sustained-release effect on the role of HCC cells.