Inhibitory effects of drug-resistant oophoroma cell line on immune activities of activated CD4+T cells and the mechanism

TIAN Jin; YAO Li

doi:10.13431/j.cnki.immunol.j.20170133

Abstract：The study aimed to determine the effects of drug-resistant oophoroma cell line SKOV3/CDDP on activated CD4+ T lymphocytes and the mechanism.Middle doses intermittent induction method was used to establish a human CDDP-resistant ovarian cancer cell line (SKOV3/CDDP).Then ELISA was employed to measure the concentration of TGF-β1 and [L-10 secreted by SKOV3/CDDP and SKOV3 cells;ELISA was also used to detect the concentration of IL-2 secreted by activated CD4 +T lymphocytes which was cultivated with normal culture medium,20％ serum of SKOV3/CDDP or 20％ serum of SKOV3 cells.The proliferation of activated CD4 + T lymphocytes was detected by counting after co-culture with SKOV3/CDDP cells.Western blot was used to test the levels of NF-κB and Snail in CD4+T lymphocytes and the levels of TGF-β and P-smad2/3 in SKOV3/CDDP cells.The adhesive ability of activated human CD4+T lymphocytes was probed with SKOV3/CDDP cells using HE staining.Data showed that the drug resistant index of SKOV3/CDDP cells were 32.4,demonstrating cross resistance to other chemotherapy drugs (ADM,PTX).Compared with SKOV3 cells,SKOV3/CDDP cells had low apoptosis rate,and secreted more TGF-β1.The supernatant of SKOV3/CDDP cells suppressed the proliferation of activated CD4+ T cells by inhibiting the secretion of IL-2.The expression of NF-κB and Snail was decreased in CD4+T lymphocytes after stimulation with the drug-resistant cell;the expression of TGF-β1 and P-smad2/3 were higher in SKOV3/CDDP cells as compared with SKOV3.Taken together,suppression of immune activities of activated CD4+ T cells by SKOV3/CDDP cells may correlate with TGF-β1,and achieve by inhibiting the NF-κB-Snail signaling pathway in CD4+ T cells.