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[ABSTRACT]AIM:ToinvestigatetheeffectoforidoninontheinvasionandmigrationofhumanlungcancerNCI-H460 cells.METHODS:NCI-H460 cells were divided into high-dose (HD), middle-dose (MD) and low-dose (LD) oridonin groups (cultured with 40, 20 and 10μmol/L of oridonin, respectively, as experimental groups), and normal (N) group ( treated without oridonin as control ) .The cell growth was observed .The cell proliferation was detected by MTT as-say.Boyden chamber was used to determine the cell invasive capacity .The cell migration was also measured .The levels of MMP-2 and MMP-9 were assayed by Western blotting .RESULTS:The cell counts in the experimental groups were lower than that in N group .The cell proliferation was inhibited as the inhibitory rates were 48.94%, 36.17%and 19.15% for HD group, MD group and LD group, respectively.The numbers of the invasive cells were 26.67 ±5.16 for HD group, 36.17 ±5.08 for MD group, and 44.33 ±5.50 for LD group.The migration rates in the experimental groups were lower than that in N group .The expression of MMP-2 and MMP-9 decreased dependent on the oridonin dose as follows: HD group
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DC group.IL-12 secretion in IL-18/fusion group was higher than that in the fusion group,and IL-12 in the pulsed DC group was higher than that in the DC group.The in vitro killing rates of the 4 groups were 79.73%,50.68%,35.81% and 4.05%,respectively.Tumor forming time in IL-18/fusion group([12.82?2.85]d) was longer than those in the pulsed DC group([8.52?1.97]d,P
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0.05).The positive rates of CD1a and CD83 in IL-18+TNF-? group were higher than those in other 2 groups.The positive rate of HLA-DR in IL-18+TNF-? group was higher than that in IL-18 group.No difference between IL-18 group and TNF-? group in the potency of stimulating T cell proliferation was found,whereas the stimulating potency in IL-18+TNF-? group was higher than that in IL-18 group and TNF-? group.IL-12 in IL-18+TNF-? group at 48 h and 72 h was higher than that in IL-18 group and TNF-? group(P
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AIM: To investigate the role of nuclear factor ?B (NF-?B) in the induction of IL-8 gene by TNF-? in colon cancer cells and the effect of antioxidant on the induction of IL-8. METHODS: ELISA was used to detect the concentrations of IL-8. IL-8 mRNA was analyzed by using RT-PCR. NF-?B in the cell nuclei was detected with electrophoretic mobility shift assay. RESULTS: (1) IL-8 production and IL-8 mRNA expression induced by TNF-? was blocked by pyrrolidine dithiocarbamate (PDTC). (2) TNF-? triggered the activation and translocation of NF-?B and PDTC inhibited the activation of NF-?B induced by TNF-?. CONCLUSION: The induction of IL-8 gene and protein by TNF-? is dependent on the activation of NF-?B. Antioxidants may inhibit the induction of IL-8 gene and protein through inhibiting NF-?B activation.
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Objective To investigate the effects of crude extracted proteins from lesions of condyloma acuminata on immunophenotypes of dendritic cells.Methods Plastic-adherent mononuclear cells(MNCs)were isolated from umbilical cord blood or peripheral blood and cultured in media containing cytokines(GM-CSF,IL-4and LPS).The morphology and phenotypes of these cells were analyzed by flow cytometry and microscopy in12day's culture.Cells on the fourth day were incubated with crude extracts from lesions of condyloma acuminata,foreskin proteins,and PBS,respectively,followed by phenotypic analysis after9-12days' culture.Results Expression of antigens CD1a,CD80,CD86,MHC-I,MHC-II,CD14,CD54was detected after12days' cul-ture.It was shown that MNCs could be induced to differentiate to mature dendritic cells in our culture system.After incubation with crude extracts from lesions of condyloma acuminata for another9-12days,expression of CD86and HLA-DR was increased on dendritic cells.Conclusions Compared to foreskin and PBS pulsed dendritic cells,expression of CD86and HLA-DR is upregulated on dentritic cells after pulsing with condyloma acuminata lesion proteins.The data suggest that crude extracts from lesions of condyloma acuminata might enhance antigen-pre-senting capacity of dendritic cells and strengthen activation of T lymphocytes.