Effect of FOXP3 expression in pancreatic carcinoma cells on the maturation and immunologic function of dendritic cells / 中华胰腺病杂志

ABSTRACT

Objective To investigate the influence of FOXP 3 expression in pancreatic carcinoma cells (PCCs) on the maturation and immunologic function of dendritic cells (DCs).Methods The siRNA sequences targeting FOXP3 gene (siRNA-FOXP3) and negative control siRNA (siRNA-NC) were specifically designed and transfected into PCCs , then the level of IL-10 and TGF-β1 of culture supernatant were detected by ELISA.The supernatants of pancreatic carcinoma cell transfected by FOXP 3-siRNA were collected , then it was mixed with GM-CSF and IL-4 to induce the differentiation of DCs .Flow cytometric analysis were used to measure the expression of surface markers CD 86 , CD80 , HLA DR on DCs which were treated with supernatants . The levels of IL-12p70, IFN-γin supernatants were detected by ELISA .The DCs were co-cultured with T lymphocytes, and then the lymphocytes proliferation and cytoxicity were analyzed by CCK -8 assays.Results Compared with PANC1 with siRNA-NC transfection, PANC1 with siRNA-FOXP3 transfection had a decreased expression of IL-10, TGF-β1 [(8.93 ±3.06)ng/L vs (26.60 ±5.57)ng/L;(2 544 ±78)ng/L vs (2 856 ± 92)ng/L], the positive expression rate of CD86, HLA DR in DCs cultured in the medium containing the supernatants of the pancreatic carcinoma cell transfected by siRNA-FOXP3 was significantly increased [(28.10 ±3.11)%vs (13.90 ±0.42)%;(66.15 ±4.17)%vs (43.15 ±3.32)%], the expression of IL-12p70, IFN-γwas significantly increased [(52.75 ±7.89)ng/L vs (26.14 ±4.50)ng/L, (898.43 ±88.82) ng/L vs (412.76 ±24.68) ng/L], after co-culture with lymphocytes at ratios of 15, 110, 120, the proliferation was significantly increased [(95.27 ±3.80)% vs (71.77 ±5.70)%, (78.97 ±5.73)% vs (52.30 ±8.72)%, (57.60 ±4.36)% vs (43.73 ±6.01)%], and the cytoxicity of CTL to PANC1 cells with 120, 140 co-culture was significantly increased [(28.44 ±5.20)% vs (8.82 ±2.29)%, (40.85 ± 5.15)% vs (17.38 ±4.86)%], and the difference between the two groups was statistically significant (P<0.05).Conclusions FOXP3 expression in PCCs can inhibit the maturation and immunologic function of DCs.