Preparation of verteporfin PLGA nanoparticles and the effect on hepatocellular cancer in mice
Objective To prepare a verteporfin-loaded poly(lactic-co-glycolic acid)nanoparticle(PLGA-VP NPs)and to verify its inhibitory effect on hepatocellular carcinoma cell line Hepa 1-6 and its in vivo safety.Methods PLGA-VP NPs were prepared by emulsion evaporation method,and the particle size and potential of PLGA-VP NPs were detected by dynamic light scattering(DLS).Mouse hepatocellular carcinoma cell line Hepa 1-6 was cultured in vitro and divided into control group(without any treatment),PLGA NPs group(adding PLGA NPs)and PLGA-VP NPs group(adding PLGA-VP NPs).The cell proliferation was detected by the Alamar Blue;the uptake of PLGA-VP NPs and apoptosis rate of cells were detected by flow cytometry;the expression levels of programmed death-ligand 1(PD-L1)protein,B-cell lymphoma-2(Bcl-2)protein and Bcl-2 associated X protein(Bax)were detected by Western blotting.C57 mice were divided into control group(phosphate buffered saline 100 μL)and PLGA-VP NPs group(tail vein injection of 2 μg·μL-1 PLGA-VP NPs 100 μL).The main organs of mice were observed by hematoxylin-eosin(HE)staining.Results The particle size of PLGA-VP NPs was about(124.00±1.80)nm,and the potential was about(-38.00±1.40)mV.When the concentrations of PLGA-VP NPs were 0,0.5,1,2,2.5,3,4,5 mg·mL-1,the cell survival rates were(100.00±2.02)％,(87.33±3.74)％,(55.86±9.71)％,(30.32±6.30)％,(26.03±2.60)％,(23.81±2.09)％,(23.63±1.29)％and(19.75±3.32)％;the half maximal inhibitory concentration of PLGA-VP NPs on Hepa 1-6 was 0.87 mg·mL-1.The uptake rates of Hepa 1-6 to PLGA-VP NPs at 0,2,4,8,12 and 24 h were 0,(3.27±0.10)％,(7.32±1.36)％,(27.03±1.11)％,(44.97±1.43)％and(68.37±0.94)％,respectively.The apoptosis rates of the control group,the PLGA NPs group and the PLGA-VP NPs group were(9.26±1.42)％,(11.08±1.16)％and(27.27±1.12)％;the relative expression levels of PD-L1 protein were 1.00±0.01,1.00±0.08 and 0.21±0.06;the relative expression levels of Bcl-2 protein were 1.20±0.02,0.43±0.01 and 0.54±0.07;the relative expression levels of Bax protein were 0.35±0.02,1.09±0.07 and 1.30±0.06,respectively.The above indexes show statistically significant differences in between the PLGA-VP NPs group and the control group,PLGA NPs group(all P＜0.05).After HE staining,comparing to the normal group,there was no obvious abnormality in the main organs of the mice in the PLGA-VP NPs group.Conclusions PLGA-VP NPs were successfully synthesized,and the nanoparticles have inhibitory effects on Hepa 1-6 hepatocellular carcinoma cells,can reduce the expression of PD-L1 protein,and have good in vivo safety.
verteporfinhepatocellular carcinomapoly(lactic-co-glycolic acid)programmed cell death 1 ligand 1
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