The ultimate goal in phosphoinositides cellular metabolism is to decipher their global functional organization and coordination of the com- partmentalized signaling processes. In this report we present evidence linkin...The ultimate goal in phosphoinositides cellular metabolism is to decipher their global functional organization and coordination of the com- partmentalized signaling processes. In this report we present evidence linking nuclear phos- phoinositides cycle with endoplasmic reticulum synthesis and function. The rapid transformation of [3H]inositol-labeled phosphoinositides in the intact nuclei (IN) was captured in chase studies for 0-5 min, followed by examination of phosphatidylinositides in the inner nuclear me- mbrane (INM), the outer nuclear membrane (ONM) and endoplasmic reticulum (ER). We revealed that synthesis of phosphatidylinositol phosphates (PIPs) occurs in ONM and the de- phosphorylation takes place in the INM. The rapid transformation of the radiolabeled PIPs in ONM reverberated in their appearance and successive transformation in INM, and in the 5min chased nuclei was tracked to ONM as the re- emerging radiolabeled phosphatidylinositol (PI). These chase-uncovered changes in ONM and INM PIPs profiles allow us to conclude that the observed conversions in the nuclear membrane continuum are induced by the lateral movement of the membrane and its transit from the cytosolic to nuclear and back to cytosolic environment. The suggested membrane synthesisinduced movement provides the means to transport the membrane- and the membrane lipid ligand-associated cytosolic proteins to the intranuclear spaces and renewal of INM. Export of the nuclear components interacting with the modified INM, by exiting from nuclear to cytosolic site, endows ER with a steady influx of the membrane that is conditioned to generate vesicles according to the nucleus delivered templates.展开更多
Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-rela...Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration（AMD）,was associated with altered activation of phosphatidylinositide 3-kinase（PI3K）,Akt,and glycogen synthase kinase 3（GSK3）.We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells（PBMC） obtained from AMD patients.In the patients＇ PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates（e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU）,indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.展开更多
Phosphatidylinositide 3-kinase （PI3K）/protein kinase B （PKB, Akt） pathway plays a major role in proliferation and survival of many types of cells. The inhibitory effect of LY294002, widely ap- plied as an inhibito...Phosphatidylinositide 3-kinase （PI3K）/protein kinase B （PKB, Akt） pathway plays a major role in proliferation and survival of many types of cells. The inhibitory effect of LY294002, widely ap- plied as an inhibitor of PI3K, in combination with gemcitabine on proliferation of PANC-1 ceils was investigated. The expression of PI3K, phosphorylated AM （p-Akt） and multidrng-resistance like protein （MRP） in normal pancreas tissues, chronic pancreatitis tissues and pancreatic carcinoma tissues was de- tected. The effects of LY294002 combined with gemcitabine on proliferation of PANC-1 cells and pro- tein levels of p-Akt and MRP were detected. The results showed that the positive expression rate of PI3K, p-Akt and MRP in pancreatic carcinoma tissues was significantly higher than that in normal pan- creas tissues and chronic pancreatitis tissues （P〈0.01 and P〈0.05 respectively）. LY294002 could effec- tively enhance the inhibitory effect of gemcitabine on proliferation of PANC-1 cells. Furthermore, Western blotting revealed that LY294002 combined with gemcitabine reduced the protein levels of p-Akt and MRP, which contributed to the inhibition of proliferation. It is concluded that LY294002 in combination with gemcitabine may represent an alternative therapy for pancreatic carcinoma.展开更多
文摘The ultimate goal in phosphoinositides cellular metabolism is to decipher their global functional organization and coordination of the com- partmentalized signaling processes. In this report we present evidence linking nuclear phos- phoinositides cycle with endoplasmic reticulum synthesis and function. The rapid transformation of [3H]inositol-labeled phosphoinositides in the intact nuclei (IN) was captured in chase studies for 0-5 min, followed by examination of phosphatidylinositides in the inner nuclear me- mbrane (INM), the outer nuclear membrane (ONM) and endoplasmic reticulum (ER). We revealed that synthesis of phosphatidylinositol phosphates (PIPs) occurs in ONM and the de- phosphorylation takes place in the INM. The rapid transformation of the radiolabeled PIPs in ONM reverberated in their appearance and successive transformation in INM, and in the 5min chased nuclei was tracked to ONM as the re- emerging radiolabeled phosphatidylinositol (PI). These chase-uncovered changes in ONM and INM PIPs profiles allow us to conclude that the observed conversions in the nuclear membrane continuum are induced by the lateral movement of the membrane and its transit from the cytosolic to nuclear and back to cytosolic environment. The suggested membrane synthesisinduced movement provides the means to transport the membrane- and the membrane lipid ligand-associated cytosolic proteins to the intranuclear spaces and renewal of INM. Export of the nuclear components interacting with the modified INM, by exiting from nuclear to cytosolic site, endows ER with a steady influx of the membrane that is conditioned to generate vesicles according to the nucleus delivered templates.
基金supported by intramural research funding of National Center for Complementary and Alternative Medicine(now is National Center for Complementary and Integrative Health),NIH,the US Department of Health and Human Services(to X.L.)and an operating grant(MOP 123279)from Canadian Institutes for Health Research(to Z.Y.)
文摘Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration（AMD）,was associated with altered activation of phosphatidylinositide 3-kinase（PI3K）,Akt,and glycogen synthase kinase 3（GSK3）.We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells（PBMC） obtained from AMD patients.In the patients＇ PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates（e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU）,indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.
文摘Phosphatidylinositide 3-kinase （PI3K）/protein kinase B （PKB, Akt） pathway plays a major role in proliferation and survival of many types of cells. The inhibitory effect of LY294002, widely ap- plied as an inhibitor of PI3K, in combination with gemcitabine on proliferation of PANC-1 ceils was investigated. The expression of PI3K, phosphorylated AM （p-Akt） and multidrng-resistance like protein （MRP） in normal pancreas tissues, chronic pancreatitis tissues and pancreatic carcinoma tissues was de- tected. The effects of LY294002 combined with gemcitabine on proliferation of PANC-1 cells and pro- tein levels of p-Akt and MRP were detected. The results showed that the positive expression rate of PI3K, p-Akt and MRP in pancreatic carcinoma tissues was significantly higher than that in normal pan- creas tissues and chronic pancreatitis tissues （P〈0.01 and P〈0.05 respectively）. LY294002 could effec- tively enhance the inhibitory effect of gemcitabine on proliferation of PANC-1 cells. Furthermore, Western blotting revealed that LY294002 combined with gemcitabine reduced the protein levels of p-Akt and MRP, which contributed to the inhibition of proliferation. It is concluded that LY294002 in combination with gemcitabine may represent an alternative therapy for pancreatic carcinoma.
