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OBJECTIVE To investigate the effect of phosphotyrosine interaction domain containing 1 (PID1, NYGGF4) onpromotion of IR and HCC, and explore its underlying mechanisms. METHODS Lentivirus were used to mediate the knockdown of PID1 in HFD induced IR mouse model as well as ob/ob mice. Intraperitoneal glucose and insulin tolerance were performed 4 weeks after lentivirus injection. Hydrodynamics-based transfection was applied to induce the liver specific overexpression of PID1. Flow cytometry was exerted to detect the proportion and function of immune cells.qRT-PCR and Western blot were used to detect the expression of downstream pathways of PID1. Liquid chromatography-mass spectrometry (LC-MS) and co-immunoprecipitation (Co-IP) were conducted to identify proteins interacting with PID1.Chromatin immunoprecipitation(ChIP)was operated to measure the modification of H3K4me3 of PID1 promoter.RESULTS PID1 restriction improved insulin resistance,hyperglycemia and fatty liver. Conversely, hepatic knockdown of PID1 attenuated liver xenografted tumor growth. Moreover,PID1 liver-specific protooncogenes via hydrodynamics-based transfection established a primary hepatocellular carcinoma mouse model,induced an immunosuppressive environment,with the reduction of CD3+,CD4+,CD8+T cells,retarded maturation of dendritic cells(DCs),pronounced differentiation of regulatory T cells(Tregs),and recruitment of MDSC.In addition,PID1 overexpression activated prolifer-ation related genes, promoted anti-inflammatory genes, suppressed pro-inflammatory genes, induced glycolysis and lipid metabolism genes to facilitate tumorigenesis in liver. Importantly, PID1 exerted its tumor-promoting function through binding to epidermal growth factor receptor(EGFR)and activation of downstream KRAS/ERK pathway.As such,PID1 exist trimethylation of histone H3 at lysine 4(H3K4me3) modification and IR up-regulated the expression of PID1 by activation the H3K4me3 modification. CONCLUSION PID1 is a new gene that exerts both liver cancer-promoting and insulin resistance inducing function.IR accelerates liver cancer development and progression partially dependent on the activation of PID1.
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OBJECTIVE To explore the immunity-enhancing effect of ABP-AW1, a low-molecular-mass polysaccharides isolated from Agaricus blazei Murill, on immunosuppressive mice. METHODS ICR mice were ip injected cyclophosphamide 80 mg · kg-1, once daily for 3 d, to establish an immuno?suppressive mouse model. Then, ABP-AW1125, 250 and 500 mg · kg-1 were ig given to the immuno?suppressive mice,respectively, once daily for 7 d. The mouse thymus index and spleen index were calcu?lated, and the phagocytic function of phagocytes was determined using carbon clearance test. Splenic lym?phocyte proliferation was measured by MTT method. The interleukin 2 (IL-2) and interferon γ (IFN-γ) production from splenic lymphocytes was examined by ELISA. The splenic lymphocyte CD4+/CD8+ratio was determined by flow cytometric analysis. RESULTS Compared with normal control group, the thymus index, spleen index and phagocytic index of phagocytes in the immunosuppressive model mice declined (P<0.05). ABP-AW1250 and 500 mg·kg-1 treatment significantly increased the thymus index, spleen index and phagocytic index in immunosuppressive mice (P<0.05). Compared with normal control group, concanavalin A (Con A) and lipopolysaccharide (LPS) induced T and B lymphocyte proliferation, respectively, and IL-2 and IFN-γproduction from splenic lymphocytes in the immunosuppressive model mice was lower (P<0.05). Compared with model group, ABP-AW1250 and 500 mg·kg-1 promoted Con A and LPS induced T and B lymphocyte proliferation (P<0.05) , and elevated IL-2 and IFN-γ production from splenic lymphocytes (P<0.05). In addition, ABP-AW1250 and 500 mg·kg-1 reversed the decreased splenocyte CD4+/CD8+ratio in immunosuppressive model mice (P<0.05). CONCLUSION ABP-AW1 has immuneity-enhancing effect on cyclophosphamide-induced immunosuppressive mice.
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OBJECTIVE To investigate the effect of phosphotyrosine interaction domain containing 1 (PID1, NYGGF4) on promotion of IR and HCC, and explore its underlying mechanisms. METHODS Lentivirus were used to mediate the knockdown of PID1 in HFD induced IR mouse model as well as ob/ob mice. Intraperitoneal glucose and insulin tolerance were performed 4 weeks after lentivirus injection. Hydrodynamics-based transfection was applied to inducethe liver specific overexpression of PID1. Flow cytometry was exerted to detect the proportion and function of immune cells. qRT-PCR and Western blot were used to detect the expression of downstream pathways of PID1.Immunoprecipitation was used to determine the receptor of PID1. Chromatin immunoprecipitation (ChIP) was operated to measure the modification of H3K4me3 of PID1 promoter. RESULTS PID1 restriction improved insulin resistance, hyperglycemia and fatty liver. Conversely, hepatic knockdown of PID1 attenuated liver xenografted tumor growth. Moreover, PID1 liver- specific protooncogenes via hydrodynamics- based transfection established a primary hepatocellular carcinoma mouse model, induced an immunosuppressive environment, with the reduction of CD3 +, CD4 +, CD8 +T cells, retarded maturation of dendritic cells (DCs), pronounced differentiation of regulatory T cells (Tregs), and recruitment of MDSC. In addition, PID1 overexpression activated proliferation related genes, promoted anti- inflammatory genes, suppressed pro-inflammatory genes, induced glycolysis and lipid metabolism genes to facilitate tumorigenesis in liver. Importantly, PID1 exerted its tumor-promoting function through binding to epidermal growth factor receptor (EGFR) and activation of downstream MAPK pathway. As such, PID1 exist trimethylation of histone H3 at lysine 4 (H3K4me3) modification and IR up-regulated the expression of PID1 by activation the H3K4me3 modification. CONCLUSION PID1 is a new gene that exerts both liver cancer-promoting and insulin resistance inducing function. IR accelerates liver cancer development and progression partially dependent on the activation of PID1.