Suppressive Effect Of Anisomycin On The Jurkat T Cells And Its Mechanism

Objactive:To study Suppressive effect of Anisomycin on the Jurkat T cells and its mechanism.Methods:The proliferation of 14 tumor cell lines from human different tissues was analyzed by MTT method.The colony formation of Jurkat T cells was observed by Inverted Microscope.The expression level of CD69 and CD25 on the surface of Jurkat T cells stimulated by Phorbol-12-myristate-13-acetate(PMA) and ionomycin(Ion) was evaluated by fluorescin-conjugated monoclonal antibody double labeling technique.The Jurkat T cell cycle was analyzed by PI staining and Flow Cytometry.The apoptosis of Jurkat T cells was analyzed by combining Annexin V-FITC with PI staining.The human interleukin 4,interleukin 10,interleukin 17(IL-4,IL- 10,IL- 17)and the Transforming Growth Factorβ(TGF-β) and Interferonγ(IFN-γ) that screted by Jurkat T cells were all analyzed by ELISA assay.The extracellular regulated protein kinase (ERK1/2)in Jurkat T cells was analyzed by Western Blotting.Result:The inhibitory rate of 14 tumor cells from different human tissues increased along with the increase of Anisomycin concentration,the highest inhibitory rate was 81%for HL-60 cells,the lowest inhibitory rate was 31.29%for HeLa cells.The CD69 and CD25 expressing level on the surface of Jurkat T cells was decreased along with the increase of Anisomycin concentration.It was also observed that the colony formation of Jurkat T cells decreased along with the increase of Anisomycin concentration.The Jurkat T cell cycle was inhibited obviously by Anisomycin before the phase of G0/G1.The apoptosis of Jurkat T cells was promoted by the dose of 40.0 ng/ml～160.0 ng/ml Anisomycin.The expressing level of IL-4,IL-10,IL-17,TGF-βand IFN-γwere was decreased along with the increase of Anisomycin concentration.The expressing level of ERK1/2 in Jurkat T cells did not change 0～6 h after the treatment of 40 ng/ml Anisomycin but the P-ERK upregulated during 0～6 h.Conelusion:Anisomycin can inhibit the proliferation of diverse tumor cells from different human tissues.Anisomycin can inhibit the activation and cell cycle entry of Jurkat T cells,and promote the apoptosis of Jurkat T cells.Anisomycin can also inhibit the secretion of immunologic cytokine that expressed by Jurkat T cells.The results suggest that the suppressive effect of Anisomycin on the Jurkat T cells may be related to inhibiting the TGF-βexpression and promoting the apoptosis of Jurkat T cells.