Study On Inhibition Effect And Mechanism Of SAHA Liposome On Ovarian Cancer Cells

ObjectiveSuberoylanilide hydroxamic acid is one of the classic histone deacetylase inhibitors,have certain antitumor activity,inhibit inhibit the liver cancer,lung cancer,stomach cancer and other tumors.The purpose of this paper was to adopt the liposome preparation technology of SAHA liposomes,with the prescription optimization and quality,to explore its effects and molecular mechanisms on ovarian cancer,laying the foundation for further application in vivo.MethodsA high performance liquid chromatography method was developed to evaluate the concentration of SAHA.SAHA liposomes were prepared by thin film dispersion ultrasonic method,its optimum evaluated by single factor study and orthogonal design.SAHA liposomes were investigated the particle size distribution and morphology,encapsulation efficiency and stability with HPLC.Two groups of ovarian carcinoma cell lines(SKOV3;HO8910)were exposed to SAHA 1~4groups(1、3、6 and 10 μM),CCk-8 method was employed to evaluate the inhibitory effects of SAHA;Ovarian cancer cell lines treated with SAHA 1 and 2 groups(3 or 6 μM),flow cytometry was performed following staining with Annexin V-FITC and PI for Cell cycle and Apoptosis assay;Reverse transcription olymerase chain reaction(RT-PCR)and western blotting were used to assess the mRNA and protein expression levels of phenotypic correlation factor;Establishment of human ovarian cancer cell line SKOV3,HO8910 in mouse xenograft tumor model and set up a blank liposome controlgroup and SAHA liposome drug group,the in vivo antitumor effect of SAHA liposome is studied through comparison of tumor bearing nude mice tumor inhibition rate.ResultsThe regression equation of concentration of SAHA is A=1.6865×104C-2321.2,(r2=0.999 8),with good linearity in concentration range of 5~160 μg·m L-1.The specificity precision and recovery conformed to the requirements of the experiment.The optimized preparation condition was: ratio of drug to lipid is 1:5;ratio of cholesterol to EPC was 1:5,and the volume of phosphate buffer aline(PBS)was 10.0 m L.The liposomes preparated by thin film dispersion ultrasonic method have almost spherical shape and high encapsulation efficiency(90 %)with average particle size was 160.8±7.5 nm and good stability.After 24 or 48 h of SAHA treatment,the OD value of SKOV3 and HO8910 showed a trend of gradually reduce(P<0.05);The apoptosis rates of SAHA 1and 2 groups were higher than control group(P<0.05).Compared with control group after 48 h of SAHA treatment,S phase and G2/M phase of SKOV3 cells increased,G0/G1 phase of HO8910 cells increased in SAHA1 and 2 groups(P<0.05);The expression levels of Cyclin B1 and Cdc2(p34)mRNA were lower than the control group in SAHA 1 and 2 group,while the expression levels of Caspase3,p21 and p53 mRNA expression level is significantly higher than control group.Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,53 protein markedly improved,and Cyclin B1,Cdc2(p34)protein decreased in SAHA 1~4 groups;Compared with the control group,the SAHA liposome was applied to the tumor bearing mice,the tumor was significantly reduced,the tumor inhibition rate was high,and the effect was well.ConclusionsSAHA liposomes may suppress growth,induce apoptosis and cause cycle arrest in ovarian carcinoma cells.The mechanism may be through the regulation of malignant phenotype related Caspase3 protein,p53,cyclin B1,Cdc2(p34)expression to promote cell histone acetylation level,play the antitumor effects in vivo or vitro.Therefore,liposome preparation as a new drug carrier system of Suberoylanilide hydroxamic acid,it has a good application prospect.