Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its g...Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its gene expression level and prognosis.Methods:The exosomes of hepatocellular carcinoma cells HepG2 and 7721 were isolated by ultracentrifugation,and the extracted exosomes were verified by transmission electron microscope, western blotting,and nanosight.The relative quantitative proteomics analysis was used to analyze the protein spectrum of HepG2 and 7721 cells in exosomes.The bioinformatics was used to collect the signal pathway,and the target protein AP2M1 was verified by qRTPCR.Gene Expression Profiling Interactive Analysis(GEPIA),OncoLnc,and The Cancer Genome Atlas(TCGA)database were used to analyze the expression level of AP2M1 and the survival rate of patients with liver cancer.Results:The extracellular exosomes of HepG2 and 7721 cells were isolated by ultracentrifugation.High-purity exosomes were obtained and verified by transmission electron microscopy,western blotting,and nanosight.There were 836 proteins co-expressed by the exosomes of HepG2 and 7721.By the analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,a total of 34 pathways that were related to liver cancer were collected,and the target membrane protein AP2M1 was screened.AP2M1 highly expressed in HepG2 cells,HepG2 and Huh7 cells' exosomes were detected by qRT-PCR(P<0.05).The data of GEPIA showed that comparing to the adjacent tissues,gene AP2M1 was highly expressed in hepatocarcinoma tissues.Kaplan-Meier survival curve analysis was performed by OncoLnc.It was found that the survival rate of patients with liver cancer and high expression of AP2M1 was significantly lower than patients with liver cancer and low expression of AP2M1(P<0.01).Conclusion:The high expression of membrane protein AP2M1 in the exosomes of hepatoma cells and the high expression of its gene in liver cancer tissues predicted the low survival rate.展开更多
BACKGROUND Gastric cancer is a prevalent malignant cancer with a high incidence and significantly affects the health of modern people globally.Cisplatin(DDP)is one of the most common and effective chemotherapies for p...BACKGROUND Gastric cancer is a prevalent malignant cancer with a high incidence and significantly affects the health of modern people globally.Cisplatin(DDP)is one of the most common and effective chemotherapies for patients with gastric cancer,but DDP resistance remains a severe clinical challenge.AIM To explore the function of M2 polarized macrophages-derived exosomal microRNA(miR)-588 in the modulation of DDP resistance of gastric cancer cells.METHODS M2 polarized macrophages were isolated and identified by specific markers using flow cytometry analysis.The exosomes from M2 macrophages were identified by transmission electron microscopy and related markers.The uptake of the PKH67-labelled M2 macrophages-derived exosomes was detected in SGC7901 cells.The function and mechanism of exosomal miR-588 from M2 macrophages in the modulation of DDP resistance of gastric cancer cells was analyzed by CCK-8 assay,apoptosis analysis,colony formation assay,Western blot analysis,qPCR analysis,and luciferase reporter assay in SGC7901 and SGC7901/DDP cells,and by tumorigenicity analysis in nude mice.RESULTS M2 polarized macrophages were isolated from mouse bone marrow stimulated with interleukin(IL)-13 and IL-4.Co-cultivation of gastric cancer cells with M2 polarized macrophages promoted DDP resistance.M2 polarized macrophagesderived exosomes could transfer in gastric cancer cells to enhance DDP resistance.Exosomal miR-588 from M2 macrophages contributed to DDP resistance of gastric cancer cells.miR-588 promoted DDP-resistant gastric cancer cell growth in vivo.miR-588 was able to target cylindromatosis(CYLD)in gastric cancer cells.The depletion of CYLD reversed miR-588 inhibition-regulated cell proliferation and apoptosis of gastric cancer cells exposed to DDP.CONCLUSION In conclusion,we uncovered that exosomal miR-588 from M2 macrophages contributes to DDP resistance of gastric cancer cells by partly targeting CYLD.miR-588 may be applied as a potential therapeutic target for the treatment of gastric cancer.展开更多
Astrocytes protect neurons by modulating neuronal function and survival.Astrocytes support neurons in several ways.They provide energy through the astrocyte-neuron lactate shuttle,protect neurons from excitotoxicity,a...Astrocytes protect neurons by modulating neuronal function and survival.Astrocytes support neurons in several ways.They provide energy through the astrocyte-neuron lactate shuttle,protect neurons from excitotoxicity,and internalize neuronal lipid droplets to degrade fatty acids for neuronal metabolic and synaptic support,as well as by their high capacity for glutamate uptake and the conversion of glutamate to glutamine.A recent reported astrocyte system for protection of dopamine neurons against the neurotoxic products of dopamine,such as aminochrome and other o-quinones,were generated under neuromelanin synthesis by oxidizing dopamine catechol structure.Astrocytes secrete glutathione transferase M2-2 through exosomes that transport this enzyme into dopaminergic neurons to protect these neurons against aminochrome neurotoxicity.The role of this new astrocyte protective mechanism in Parkinson´s disease is discussed.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81760612)Guangxi Key Research and Development Project(No.AB16380351)
文摘Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its gene expression level and prognosis.Methods:The exosomes of hepatocellular carcinoma cells HepG2 and 7721 were isolated by ultracentrifugation,and the extracted exosomes were verified by transmission electron microscope, western blotting,and nanosight.The relative quantitative proteomics analysis was used to analyze the protein spectrum of HepG2 and 7721 cells in exosomes.The bioinformatics was used to collect the signal pathway,and the target protein AP2M1 was verified by qRTPCR.Gene Expression Profiling Interactive Analysis(GEPIA),OncoLnc,and The Cancer Genome Atlas(TCGA)database were used to analyze the expression level of AP2M1 and the survival rate of patients with liver cancer.Results:The extracellular exosomes of HepG2 and 7721 cells were isolated by ultracentrifugation.High-purity exosomes were obtained and verified by transmission electron microscopy,western blotting,and nanosight.There were 836 proteins co-expressed by the exosomes of HepG2 and 7721.By the analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,a total of 34 pathways that were related to liver cancer were collected,and the target membrane protein AP2M1 was screened.AP2M1 highly expressed in HepG2 cells,HepG2 and Huh7 cells' exosomes were detected by qRT-PCR(P<0.05).The data of GEPIA showed that comparing to the adjacent tissues,gene AP2M1 was highly expressed in hepatocarcinoma tissues.Kaplan-Meier survival curve analysis was performed by OncoLnc.It was found that the survival rate of patients with liver cancer and high expression of AP2M1 was significantly lower than patients with liver cancer and low expression of AP2M1(P<0.01).Conclusion:The high expression of membrane protein AP2M1 in the exosomes of hepatoma cells and the high expression of its gene in liver cancer tissues predicted the low survival rate.
文摘BACKGROUND Gastric cancer is a prevalent malignant cancer with a high incidence and significantly affects the health of modern people globally.Cisplatin(DDP)is one of the most common and effective chemotherapies for patients with gastric cancer,but DDP resistance remains a severe clinical challenge.AIM To explore the function of M2 polarized macrophages-derived exosomal microRNA(miR)-588 in the modulation of DDP resistance of gastric cancer cells.METHODS M2 polarized macrophages were isolated and identified by specific markers using flow cytometry analysis.The exosomes from M2 macrophages were identified by transmission electron microscopy and related markers.The uptake of the PKH67-labelled M2 macrophages-derived exosomes was detected in SGC7901 cells.The function and mechanism of exosomal miR-588 from M2 macrophages in the modulation of DDP resistance of gastric cancer cells was analyzed by CCK-8 assay,apoptosis analysis,colony formation assay,Western blot analysis,qPCR analysis,and luciferase reporter assay in SGC7901 and SGC7901/DDP cells,and by tumorigenicity analysis in nude mice.RESULTS M2 polarized macrophages were isolated from mouse bone marrow stimulated with interleukin(IL)-13 and IL-4.Co-cultivation of gastric cancer cells with M2 polarized macrophages promoted DDP resistance.M2 polarized macrophagesderived exosomes could transfer in gastric cancer cells to enhance DDP resistance.Exosomal miR-588 from M2 macrophages contributed to DDP resistance of gastric cancer cells.miR-588 promoted DDP-resistant gastric cancer cell growth in vivo.miR-588 was able to target cylindromatosis(CYLD)in gastric cancer cells.The depletion of CYLD reversed miR-588 inhibition-regulated cell proliferation and apoptosis of gastric cancer cells exposed to DDP.CONCLUSION In conclusion,we uncovered that exosomal miR-588 from M2 macrophages contributes to DDP resistance of gastric cancer cells by partly targeting CYLD.miR-588 may be applied as a potential therapeutic target for the treatment of gastric cancer.
基金supported by ANID-FONDECYT 1170033(to JSA)ANID-STINT-CONICYT CS2018-7940(to JSA,IN,JI,MV)Swedish Research Council grant 2015-04222 to BM.
文摘Astrocytes protect neurons by modulating neuronal function and survival.Astrocytes support neurons in several ways.They provide energy through the astrocyte-neuron lactate shuttle,protect neurons from excitotoxicity,and internalize neuronal lipid droplets to degrade fatty acids for neuronal metabolic and synaptic support,as well as by their high capacity for glutamate uptake and the conversion of glutamate to glutamine.A recent reported astrocyte system for protection of dopamine neurons against the neurotoxic products of dopamine,such as aminochrome and other o-quinones,were generated under neuromelanin synthesis by oxidizing dopamine catechol structure.Astrocytes secrete glutathione transferase M2-2 through exosomes that transport this enzyme into dopaminergic neurons to protect these neurons against aminochrome neurotoxicity.The role of this new astrocyte protective mechanism in Parkinson´s disease is discussed.
