ABSTRACT
Objective:To determine the effects of Foxp3-overexpressing lung cancer cells on activated CD4+T lymphocyte.Methods: Stable Foxp3-overexpressing lung cancer cells NCIH-1299,NCIH-hFoxp3,was generated by transfection of NCIH-1299 cells with plasmid pcDNA3-hFoxp3 mediated by Lipofectamine 2000 and by selection with G418,and validated by quantitative PCR and Western blot.The expression levels of IL-8 and IL-10 secreted by NCIH-hFoxp3 and NCIH-control were measured by ELISA.IL-2 secrection by activated human CD4+T lymphocyte which was tested after stimulation with 20% conditioned medium of NCIH-hFoxp3 and NCIH-control cells.The proliferation of activated human CD4+ T lymphocytes was assessed by MTT after coculture with NCIH-hFoxp3 cells.The adhesive ability of activated human CD4+ T lymphocytes was probed with NCIH-hFoxp3 cells by immunocytochemistry.Results: Compared with NCIH-control cells,NCIH-hFoxp3 secreted high level of IL-10 and low level of IL-8.NCIH-hFoxp3 with Foxp3 overexpression significantly suppressed the proliferation,adhesive potential and IL-2 expression by activated CD4+ T cells.Conclusion: Suppression of immune activities of activated CD4+ T cells by Foxp3 overexpression in lung cancer cells may correlate with cytokine IL-8 and IL-10,which can contribute lung cancer progression.
ABSTRACT
Objective: To investigate the flavonoids from the fruits of Nicandra physaloides and clarify the immune activities of the isolated compounds. Methods: The separations and purifications were taken by silica gel and ODS chromatogram columns as well as preparative HPLC, and the structural identification based on physicochemical property, 1H-NMR, and 13C-NMR as well as HR-MS data. And the immune activities were evaluated by mice splenic lymphocyte proliferation model induced by LPS. Results: Thirteen compounds were obtained from the butanol fraction of the 70% ethanol extract of N. physaloides fruits, which were identified as 3''-hydroxy-puerarin (1), puerarin-6″-O-glucoside (2), daidzein 8-C-[α-D-apiofranosyl-(1→6)]-β-D-glucopyranoside (3), puerarin (4), 3''-methoxyl-puerarin (5), chrysin 6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (6), oroxylin A-7-O-β-D-glucuronide methyl ester (7), daidzin (8), baicalin (9), quercetin-3-O-β-D-galactopyranoside (10), quercetin 3-rutinoside (11), kaempferol 3-rutinoside (12), and quercetin 3-glucoside (13), respectively. The results indicated that compounds 1-3, 7, 9-10, and 12-13 could inhibit the proliferation of mice spleen lymphocytes and 6 could promote the proliferation in the influence of mice spleen lymphocytes proliferation induced by LPS. Conclusion: Compounds 1-7 are isolated from the plants of Solanaceae for the first time, compounds 8-13 are obtained from the plants of Nicandra Adans. firstly. And the compounds 1-3, 6, 7, 9-10, and 12-13 possibly possess the potential immune activities.