Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We em...Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We employed TWS119 to inactivate GSK-3β by phosphorylating Ser9 and explored its effect on breast cancer and NK cells. The expression of GSK-3β, natural killer group 2 member D(NKG2D) ligands, eIF2B was quantified by PCR and Western blot. We measured intracellular reactive oxygen species(ROS) and mitochondrial ROS using DCFH-DA and MitoSOX^(TM) probe,respectively, and conducted quantitative analysis of cellular respiration on 4T1 cells with mitochondrial respiratory chain complex Ⅰ/Ⅲ kits.Results: Our investigation revealed that TWS119 downregulated NKG2D ligands(H60 a and Rae1), suppressed the cytotoxicity of NK cells, and promoted the migration of 4T1 murine breast cancer cells. Nevertheless, LY290042, which attenuates p-GSK-3β formation by inhibiting the PI3K/Akt pathway, reversed these effects. We also found that higher expression of p Ser9-GSK-3β induced higher levels of ROS, and observed that abnormality of mitochondrial respiratory chain complex Ⅰ/Ⅲ function induced the dysfunction of GSK-3β-induced electron transport chain, naturally disturbing the ROS level. In addition, the expression of NOX3 and NOX4 was significantly up-regulated, which affected the generation of ROS and associated with the metastasis of breast cancer. Furthermore, we found that the expression of pSer535-eIF2B promoted the expression of NKG2D ligands(Mult-1 and Rae1) following by expression of pSer9-GSK-3β and generation of ROS.Conclusions: The PI3K/Akt/GSK-3β/ROS/eIF2B pathway could regulate NK cell activity and sensitivity of tumor cells to NK cells,which resulted in breast cancer growth and lung metastasis. Thus, GSK-3β is a promising target of anti-tumor therapy.展开更多
We characterized the murine NK cell subsets of the tumor microenvironment(TME)with low expressions of CD16 and NKG2D and investigated the chemokines that deter CD16~low NKG2D^low subsets.Our results demonstrated the a...We characterized the murine NK cell subsets of the tumor microenvironment(TME)with low expressions of CD16 and NKG2D and investigated the chemokines that deter CD16~low NKG2D^low subsets.Our results demonstrated the activation of primary and KY-1 NK cell by ligands and found that exogenous CXCL10/interferon-gamma-induced protein 10(IP-10)and fractalkine(FKN)can up-regulate the expression of CD16 and NKG2D.Moreover,both IP-10 and FKN are shown to facilitate migration,adhesion and cytotoxicity of NK cell subsets of the TME,due to the up-regulated CD16 and NKG2D.Overall,our data provide a new path by which to enhance murine NK cell cytotoxic potential and improve the quality of NK cells of the TME.展开更多
基金supported by grants from the National Natural Science Foundation of China (Grant No. 8117975 and 31770968)Tianjin Institutes for Basic Sciences (Grant No. 15JCYBJC26900 and 16JCQNJC11700)
文摘Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We employed TWS119 to inactivate GSK-3β by phosphorylating Ser9 and explored its effect on breast cancer and NK cells. The expression of GSK-3β, natural killer group 2 member D(NKG2D) ligands, eIF2B was quantified by PCR and Western blot. We measured intracellular reactive oxygen species(ROS) and mitochondrial ROS using DCFH-DA and MitoSOX^(TM) probe,respectively, and conducted quantitative analysis of cellular respiration on 4T1 cells with mitochondrial respiratory chain complex Ⅰ/Ⅲ kits.Results: Our investigation revealed that TWS119 downregulated NKG2D ligands(H60 a and Rae1), suppressed the cytotoxicity of NK cells, and promoted the migration of 4T1 murine breast cancer cells. Nevertheless, LY290042, which attenuates p-GSK-3β formation by inhibiting the PI3K/Akt pathway, reversed these effects. We also found that higher expression of p Ser9-GSK-3β induced higher levels of ROS, and observed that abnormality of mitochondrial respiratory chain complex Ⅰ/Ⅲ function induced the dysfunction of GSK-3β-induced electron transport chain, naturally disturbing the ROS level. In addition, the expression of NOX3 and NOX4 was significantly up-regulated, which affected the generation of ROS and associated with the metastasis of breast cancer. Furthermore, we found that the expression of pSer535-eIF2B promoted the expression of NKG2D ligands(Mult-1 and Rae1) following by expression of pSer9-GSK-3β and generation of ROS.Conclusions: The PI3K/Akt/GSK-3β/ROS/eIF2B pathway could regulate NK cell activity and sensitivity of tumor cells to NK cells,which resulted in breast cancer growth and lung metastasis. Thus, GSK-3β is a promising target of anti-tumor therapy.
基金the National Natural Science Foundation of China (81171975)the Tianjin Institutes for Basic Sciences (15JCYBJC26900)
文摘We characterized the murine NK cell subsets of the tumor microenvironment(TME)with low expressions of CD16 and NKG2D and investigated the chemokines that deter CD16~low NKG2D^low subsets.Our results demonstrated the activation of primary and KY-1 NK cell by ligands and found that exogenous CXCL10/interferon-gamma-induced protein 10(IP-10)and fractalkine(FKN)can up-regulate the expression of CD16 and NKG2D.Moreover,both IP-10 and FKN are shown to facilitate migration,adhesion and cytotoxicity of NK cell subsets of the TME,due to the up-regulated CD16 and NKG2D.Overall,our data provide a new path by which to enhance murine NK cell cytotoxic potential and improve the quality of NK cells of the TME.