Objective: To study the effect of anti-aging Klotho protein on immune escape mediated by regulatory T cells (Treg)/helper T cells 17 (TH17) in mice bearing cervical cancer and its mechanism. Methods: The model of cervical cancer-bearing mice were established, and the control group (normal mice), model group (cervical cancer-bearing mice model), and Klotho treatment group (cervical cancer-bearing mice treated with Klotho protein, 200 ng/d) were set up. The weight of cervical cancer tumors in mice of each group was weighed at 7 and 14 days after treatment respectively, PBMCs were separated at the same time. Flow cytometry was used to detect the changes of T lymphocyte function and the proportion of Treg and TH17 cells in mice. qPCR was used to detect the expressions of Foxp3 and RORγt, the key transcription factors of Treg/TH17 cells, in PBMCs of mice in each group. The changes of IL-17, IL-6, IL10, TGF-β and IL-23 in PBMCs were detected by ELISA. The protein expressions of Klotho, TGF-β, Foxp3 and RORγt in PBMCs of mice were detected by WB assay. Results: On the 14th day, the tumor inhibition rate of the cervical cancer-bearing mice in the Klotho group was significantly higher than that in the Model group [(52.16±8.25)% vs (23.33±6.29)% the model group to be supplemented, P< 0.05). Compared with the Control group, the ratios of Treg and TH17 cells in the lymphocytes of the tumor-bearing mice significantly increased (all P<0.05), the ratios of total T lymphocytes (CD3+), auxiliary/induced T lymphocytes (CD3+CD4+) and immune index (CD3+CD4+/CD3+CD8+ cells) decreased significantly (all P<0.05); in addition, the mRNAexpressions of Foxp3 and RORγt genes, cytokines of IL-17, IL-6, IL-10, TGF-β and IL-23, as well as protein expressions of TGF-β1, Foxp3 and RORγt increased significantly (all P <0.05), while the level of Klotho protein significantly decreased in Model group (P<0.05). Compared with the Model group, the above indicators showed opposite changes in Klotho group (P<0.05), but there was no significant difference with the Control group (all P> 0.05). Conclusion: Klotho protein may inhibit Treg/TH17 cell-mediated immune evasion in cervical cancer-bearing mice by inhibiting TGF-β1/Foxp3/RORγt signaling pathway and exert anti-tumor effect.

Objective To investigate the change and effect of SSeCKS(src suppressed c kinase substrates)in the activation of hepatic stellate cells(HSCs).Methods HSCs were isolated from normal rats,the change of SSeCKS mRNA expression on HSCs culture in vitro was determined using real.time PCR.protein level was determined by Western blot and immunofluorescence methods.A rat model of liver fibrosis was established.The expression and location of SSeCKS and α-SMA(α-smooth muscle actin)in liver tissues were detected by immunofluorescence methods.Results SSeCKS mRNA expression WaS loW in freshly isolated HSCs cell and the expression increased in activated HSCs in vitro.In liver fibrosis tissue,the number of SSeCKS-positive cells was increased and these cells were distributed along the sinusoids which also contained α-SMA positive cells.Conclusion The expression of SSeCKS was increased in activated HSCs in vitro.Therefore.SSeCKS may be involved in the liver inflammation and fibrosis.