We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repet...We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.展开更多
目的:探讨长基因间非编码RNA 00519(long intergene non-coding RNA 00519,LINC00519)调控miR-876-3p/高迁移率家族蛋白A1(high mobility group protein A1,HMGA1)轴在胃癌HGC-27细胞的增殖、凋亡、迁移和侵袭中的作用。方法:采用qPCR...目的:探讨长基因间非编码RNA 00519(long intergene non-coding RNA 00519,LINC00519)调控miR-876-3p/高迁移率家族蛋白A1(high mobility group protein A1,HMGA1)轴在胃癌HGC-27细胞的增殖、凋亡、迁移和侵袭中的作用。方法:采用qPCR检测胃癌细胞HGC-27和胃黏膜上皮细胞GES-1中LINC00519的表达水平。将HGC-27细胞按转染处理分为si-NC、si-LINC00519、si-LINC00519+anti-miR-NC和si-LINC00519+anti-miR-876-3p组,采用集落形成实验检测细胞克隆形成能力,流式细胞术检测细胞凋亡和周期分布,Transwell实验检测细胞迁移和侵袭。双荧光素酶报告实验和qPCR验证LINC00519与miR-876-3p、miR-876-3p与HMGA1之间的相互作用。结果:HGC-27细胞中LINC00519表达较GES-1细胞显著升高(P<0.05),转染siRNA后si-LINC00519组HGC-27细胞中LINC00519的表达水平较si-NC组显著降低(t=47.294,P<0.01)。与si-NC组比较,si-LINC00519组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例均显著降低(均P<0.01),凋亡率、G0/G1期细胞比例均显著升高(均P<0.01)。与si-LINC00519+anti-miR-NC组比较,si-LINC00519+anti-miR-876-3p组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例升高(均P<0.01),凋亡率、G0/G1期细胞比例显著降低(均P<0.01)。LINC00519能够靶向负调控miR-876-3p的表达,miR-876-3p靶向负调控HMGA1的表达。结论:敲降LINC00519能够通过调控miR-876-3p/HMGA1轴抑制胃癌HGC-27细胞的增殖、迁移和侵袭,诱导细胞凋亡。展开更多
Objective: Tumor metastasis is a complex, multistep process that depends on tumor cells and their communication with the tumor microenvironment. A p53 gain-of-function mutant has been shown to enhance the tumorigenesi...Objective: Tumor metastasis is a complex, multistep process that depends on tumor cells and their communication with the tumor microenvironment. A p53 gain-of-function mutant has been shown to enhance the tumorigenesis, invasion, and metastasis abilities of tumor cells. This study aimed to investigate the roles of p53 R273 H mutation in the tumor microenvironment.Methods: The in vitro and in vivo effects of the p53 R273 H mutant on the invasion and metastasis of HCT116 cells were investigated. Exosomes from wild-type and HCT116-TP53(R273 H) cells were cocultured with mouse embryonic fibroblasts(MEFs). The roles of differentially expressed exosomal micro RNAs identified by microarray analysis were investigated. The functions of the p53 R273 H mutant in tumor cells were also investigated via gene expression microarray and quantitative polymerase chain reaction(q PCR) analyses.Results: Introducing p53 R273 H mutant into HCT116 cells significantly potentiated pulmonary metastasis in vivo. In the presence of exosomes derived from HCT116-TP53(R273 H) cells, the exosomes were taken up by MEFs and became activated. Microarray analysis showed that the p53 R273 H mutation increased the exosomal levels of mi R-21-3 p and mi R-769-3 p. Intriguingly, in clinical samples, mi R-21-3 p and mi R-769-3 p levels were significantly higher in patients with a p53 mutation than in those without this mutation. Furthermore, both mi R-21-3 p and mi R-769-3 p activated fibroblasts and exerted a synergistic effect via their target genes on the transforming growth factor-β(TGF-β)/Smad signaling pathway. The activated fibroblasts excreted cytokine TGF-β and may have reciprocally induced cancer cells to undergo epithelial-mesenchymal transition(EMT). Indeed, HCT116-TP53(R273 H) cells showed increased expression of ZEB1 and SNAI2 and decreased transcription of several cell adhesion molecules.Conclusions: The mutant p53-exosomal mi R-21-3 p/mi R-769-3 p-fibroblast-cytokine circuit appears to be responsible for communication between tumor and stromal cells, with exosomal mi RNAs acting as a bridge. mi R-21-3 p and mi R-769-3 p are potential predictive markers of pulmonary metastasis and candidate targets for therapeutic interventions.展开更多
BACKGROUND Highly upregulated in liver cancer (HULC) is a long non-coding RNA (lncRNA) which has recently been identified as a key regulator in hepatocellular carcinoma (HCC) progression. However, its role in the secr...BACKGROUND Highly upregulated in liver cancer (HULC) is a long non-coding RNA (lncRNA) which has recently been identified as a key regulator in hepatocellular carcinoma (HCC) progression. However, its role in the secretion of exosomes from HCC cells remains unknown. AIM To explore the mechanism by which HULC promotes the secretion of exosomes from HCC cells. METHODS Serum and liver tissue samples were collected from 30 patients with HCC who had not received chemotherapy, radiotherapy, or immunotherapy before surgery. HULC expression in serum exosomes and liver cancer tissues of patients was measured, and compared with the data obtained from healthy controls and tumor adjacent tissues. The effect of HULC upregulation in HCC cell lines and the relationship between HULC and other RNAs were studied using qPCR and dualluciferase reporter assays. Nanoparticle tracking analysis was performed to detect the quantity of exosomes.RESULTS HULC expression in serum exosomes of patients with HCC was higher than that in serum exosomes of healthy controls, and HULC levels were higher in liver cancer tissues than in tumor adjacent tissues. The expression of HULC in serum exosomes and liver cancer tissues correlated with the tumor-node-metastasis (TNM) classification, and HULC expression in tissues correlated with that in serum exosomes. Upregulation of HULC promoted HCC cell growth and invasion and repressed apoptosis. Notably, it also facilitated the secretion of exosomes from HCC cells. Moreover, qPCR assays showed that HULC repressed microRNA-372-3p (miR-372-3p) expression. We also identified Rab11a as a downstream target of miR-372-3p. Dual-luciferase reporter assays suggested that miR-372-3p could directly bind both HULC and Rab11a. CONCLUSION Our findings illustrate the importance of the HULC/miR-372-3p/Rab11a axis in HCC and provide new insights into the molecular mechanism regulating the secretion of exosomes from HCC cells.展开更多
Background:Cell competition is an important feature in pancreatic cancer(PC)progression,but the underlying mechanism remains elusive.This study aims to explore the role of exosomes derived from normal pancreatic ducta...Background:Cell competition is an important feature in pancreatic cancer(PC)progression,but the underlying mechanism remains elusive.This study aims to explore the role of exosomes derived from normal pancreatic ductal epithelial cells involved in PC progression.Methods:PC cells and pancreatic stellate cells(PSCs)were treated with exosomes isolated from pancreatic ductal epithelial cells.Cell proliferation was assessed by CCK8 assays.Cell migration and invasion were assessed by Transwell assays.PC and matched adjacent non-tumor tissue specimens were obtained from 46 patients pathologically diagnosed with PC at Peking University First Hospital from 2013 to 2017.Tissue miR-485-3p and p21-activated kinase-1(PAK1)expression was examined by real-time polymerase chain reaction(RT-PCR),and the relationship of the two was analyzed using Pearman’s product-moment correlation.The clinical significance of miR-485-3p was analyzed using the Chi-square test,Wilcoxon rank-sum test,and Fisher exact probability,respectively.The binding of miR-485-3p to PAK15’-untranslated region(5’-UTR)was examined by luciferase assay.PC cells were xenografted into nude mice as a PC metastasis model.Results:Exosomes from pancreatic ductal epithelial cells suppressed PC cell migration and invasion as well as the secretion and migration of PSCs.MiR-485-3p was enriched in the exosomes of pancreatic ductal epithelial cells but deficient in those of PC cells and PSCs,in accordance with the lower level in PSCs and PC cells than that in pancreatic ductal cells.And the mature miR-485-3p could be delivered into these cells by the exosomes secreted by normal pancreatic duct cells,to inhibit PC cell migration and invasion.Clinical data analysis showed that miR-485-3p was significantly decreased in PC tissues(P<0.05)and was negatively associated with lymphovascular invasion(P=0.044).As a direct target of miR-485-3p,PAK1 was found to exert an inhibitory effect on PC cells,and there was a significantly negative correlation between the expression levels of miR-485-3p and PAK1(r=-0.6525,P<0.0001)in PC tissues.Moreover,miR-485-3p could suppress PC metastasisin vivo by targeting p21-activated kinase-1.Conclusions:Exosomal miR-485-3p delivered by normal pancreatic ductal epithelial cells into PC cells inhibits PC metastasis by directly targeting PAK1.The restoration of miR-485-3p by exosomes or some other vehicle might be a novel approach for PC treatment.展开更多
Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI....Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI.However,the mechanisms underlying the regulatory effect of autophagy in AKI among patients undergoing cardiac surgeries are poorly understood.In this study,we aimed to test the hypothesis that exosomal microRNAs(miRNAs)regulate autophagy in tubular epithelial cells after AKI.Methods:Plasma exosomal RNA was extracted from young and elderly AKI patients undergoing cardiac surgery,and the miRNAs expression during the perioperative period were analyzed using next-generation sequencing.The screened miRNAs and their target genes were subjected to gene oncology function and Kyoto Encyclopedia of Genes and Genome enrichment analyses.Renal tubular epithelial cell line(HK-2 cells)was cultured and hypoxia/reoxygenation(H/R)model was established,which is an in vitro renal ischemia/reperfusion(I/R)model.We used Western blot analysis,cell viability assay,transfection,luciferase assay to investigate the mechanisms underlying the observed increases in the levels of renal I/R injury-mediated exosomal miRNAs and their roles in regulating HK-2 cells autophagy.Results:miR-590-3p was highly enriched in the plasma exosomes of young AKI patients after cardiac surgery.Increased levels of miR-590-3p led to the increases in the expression of autophagy marker proteins,including Beclin-1 and microtubule associated protein 1 light chain 3 beta(LC3II),and prolonged the autophagic response in HK-2 cells after H/R treatment.These effects were achieved mainly via increases in the exosomal miR-590-3p levels,and the tumor necrosis factor receptor-associated factor 6 protein was shown to play a key role in I/R injury-mediated autophagy induction.Conclusion:Exosomes released from HK-2 cells after renal I/R injury regulate autophagy by transferring miR-590-3p in a paracrine manner,which suggests that increasing the miR-590-3p levels in HK-2 cell-derived exosomes may increase autophagy and protect against kidney injury after renal I/R injury.展开更多
Glioma-associated microglial cells,a key component of the tumor microenvironment,play an important role in glioma progression.In this study,the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were c...Glioma-associated microglial cells,a key component of the tumor microenvironment,play an important role in glioma progression.In this study,the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were chosen.First,circadian gene expression in glioma cells co-cultured with either M1 or M2 microglia was assessed and the exosomes of M2-polarized and unpolarized BV-2 microglia were extracted.Subsequently,we labeled the exosomes with PKH67 and treated GL261 cells with them to investigate the exosome distribution.GL261 cell phenotypes and related protein expression were used to explore the role of M2 microglial exosomes in gliomas.Then a specific miR-7239-3p inhibitor was added to verify miR-7239-3p functions.Finally,the mouse subcutaneous tumorigenic model was used to verify the tumorigenic effect of M2 microglial exosomes in vivo.Our results showed that in gliomas co-cultured with M2 microglia,the expression of the BMAL1 protein was decreased(P<0.01),while the expression of the CLOCK protein was increased(P<0.05);opposite results were obtained in gliomas co-cultured with M1 microglia.After treatment with M2 microglial exosomes,the apoptosis of GL261 cells decreased(P<0.001),while the viability,proliferation,and migration of GL261 cells increased.Increased expression of N-cadherin and Vimentin,and decreased E-cadherin expression occurred upon treatment with M2 microglial exosomes.Addition of an miR-7239-3p inhibitor to M2 microglial exosomes reversed these results.In summary,we found that miR-7239-3p in the glioma microenvironment is recruited to glioma cells by exosomes and inhibits Bmal1 expression.M2 microglial exosomes promote the proliferation and migration of gliomas by regulating tumor-related protein expression and reducing apoptosis.展开更多
基金supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund,No.22HHXBSS00047(to PL)the National Natural Science Foundation of China,Nos.82072166(to PL),82071394(to XG)+4 种基金Science and Technology Planning Project of Tianjin,No.20YFZCSY00030(to PL)Science and Technology Project of Tianjin Municipal Health Commission,No.TJWJ2021QN005(to XG)Tianjin Key Medical Discipline(Specialty)Construction Project,No.TJYXZDXK-006ATianjin Municipal Education Commission Scientific Research Program Project,No.2020KJ164(to JZ)China Postdoctoral Science Foundation,No.2022M712392(to ZY).
文摘We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.
文摘目的:探讨长基因间非编码RNA 00519(long intergene non-coding RNA 00519,LINC00519)调控miR-876-3p/高迁移率家族蛋白A1(high mobility group protein A1,HMGA1)轴在胃癌HGC-27细胞的增殖、凋亡、迁移和侵袭中的作用。方法:采用qPCR检测胃癌细胞HGC-27和胃黏膜上皮细胞GES-1中LINC00519的表达水平。将HGC-27细胞按转染处理分为si-NC、si-LINC00519、si-LINC00519+anti-miR-NC和si-LINC00519+anti-miR-876-3p组,采用集落形成实验检测细胞克隆形成能力,流式细胞术检测细胞凋亡和周期分布,Transwell实验检测细胞迁移和侵袭。双荧光素酶报告实验和qPCR验证LINC00519与miR-876-3p、miR-876-3p与HMGA1之间的相互作用。结果:HGC-27细胞中LINC00519表达较GES-1细胞显著升高(P<0.05),转染siRNA后si-LINC00519组HGC-27细胞中LINC00519的表达水平较si-NC组显著降低(t=47.294,P<0.01)。与si-NC组比较,si-LINC00519组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例均显著降低(均P<0.01),凋亡率、G0/G1期细胞比例均显著升高(均P<0.01)。与si-LINC00519+anti-miR-NC组比较,si-LINC00519+anti-miR-876-3p组HGC-27细胞克隆数、迁移侵袭数、S期细胞比例升高(均P<0.01),凋亡率、G0/G1期细胞比例显著降低(均P<0.01)。LINC00519能够靶向负调控miR-876-3p的表达,miR-876-3p靶向负调控HMGA1的表达。结论:敲降LINC00519能够通过调控miR-876-3p/HMGA1轴抑制胃癌HGC-27细胞的增殖、迁移和侵袭,诱导细胞凋亡。
基金supported by grants from the National Key R & D Program of China (No. 2017YFC0906601, No. 2017ZX10203205-003-001 and No. 2016YFC0901403)the National Natural Science Foundation (No. 81572840, No. 81572365, No. 81728015 and No. 81872033)+1 种基金the Nonprofit Central Research Institute Fund of CAMS (No. 2018RC310011)the CAMS Innovation Fund for Medical Sciences (No. 2016-I2M-1-001, No. 2017-I2M-3005 and No. 2019-I2M-1-003) in China
文摘Objective: Tumor metastasis is a complex, multistep process that depends on tumor cells and their communication with the tumor microenvironment. A p53 gain-of-function mutant has been shown to enhance the tumorigenesis, invasion, and metastasis abilities of tumor cells. This study aimed to investigate the roles of p53 R273 H mutation in the tumor microenvironment.Methods: The in vitro and in vivo effects of the p53 R273 H mutant on the invasion and metastasis of HCT116 cells were investigated. Exosomes from wild-type and HCT116-TP53(R273 H) cells were cocultured with mouse embryonic fibroblasts(MEFs). The roles of differentially expressed exosomal micro RNAs identified by microarray analysis were investigated. The functions of the p53 R273 H mutant in tumor cells were also investigated via gene expression microarray and quantitative polymerase chain reaction(q PCR) analyses.Results: Introducing p53 R273 H mutant into HCT116 cells significantly potentiated pulmonary metastasis in vivo. In the presence of exosomes derived from HCT116-TP53(R273 H) cells, the exosomes were taken up by MEFs and became activated. Microarray analysis showed that the p53 R273 H mutation increased the exosomal levels of mi R-21-3 p and mi R-769-3 p. Intriguingly, in clinical samples, mi R-21-3 p and mi R-769-3 p levels were significantly higher in patients with a p53 mutation than in those without this mutation. Furthermore, both mi R-21-3 p and mi R-769-3 p activated fibroblasts and exerted a synergistic effect via their target genes on the transforming growth factor-β(TGF-β)/Smad signaling pathway. The activated fibroblasts excreted cytokine TGF-β and may have reciprocally induced cancer cells to undergo epithelial-mesenchymal transition(EMT). Indeed, HCT116-TP53(R273 H) cells showed increased expression of ZEB1 and SNAI2 and decreased transcription of several cell adhesion molecules.Conclusions: The mutant p53-exosomal mi R-21-3 p/mi R-769-3 p-fibroblast-cytokine circuit appears to be responsible for communication between tumor and stromal cells, with exosomal mi RNAs acting as a bridge. mi R-21-3 p and mi R-769-3 p are potential predictive markers of pulmonary metastasis and candidate targets for therapeutic interventions.
基金Supported by Tianjin Clinical Research Center for Organ Transplantation Project,No.15ZXLCSY00070The Nonprofit Central Research Institute Fund of Chinese Academy of Medical Sciences,No.2018PT32021
文摘BACKGROUND Highly upregulated in liver cancer (HULC) is a long non-coding RNA (lncRNA) which has recently been identified as a key regulator in hepatocellular carcinoma (HCC) progression. However, its role in the secretion of exosomes from HCC cells remains unknown. AIM To explore the mechanism by which HULC promotes the secretion of exosomes from HCC cells. METHODS Serum and liver tissue samples were collected from 30 patients with HCC who had not received chemotherapy, radiotherapy, or immunotherapy before surgery. HULC expression in serum exosomes and liver cancer tissues of patients was measured, and compared with the data obtained from healthy controls and tumor adjacent tissues. The effect of HULC upregulation in HCC cell lines and the relationship between HULC and other RNAs were studied using qPCR and dualluciferase reporter assays. Nanoparticle tracking analysis was performed to detect the quantity of exosomes.RESULTS HULC expression in serum exosomes of patients with HCC was higher than that in serum exosomes of healthy controls, and HULC levels were higher in liver cancer tissues than in tumor adjacent tissues. The expression of HULC in serum exosomes and liver cancer tissues correlated with the tumor-node-metastasis (TNM) classification, and HULC expression in tissues correlated with that in serum exosomes. Upregulation of HULC promoted HCC cell growth and invasion and repressed apoptosis. Notably, it also facilitated the secretion of exosomes from HCC cells. Moreover, qPCR assays showed that HULC repressed microRNA-372-3p (miR-372-3p) expression. We also identified Rab11a as a downstream target of miR-372-3p. Dual-luciferase reporter assays suggested that miR-372-3p could directly bind both HULC and Rab11a. CONCLUSION Our findings illustrate the importance of the HULC/miR-372-3p/Rab11a axis in HCC and provide new insights into the molecular mechanism regulating the secretion of exosomes from HCC cells.
基金National Natural Science Foundation of China(No.82171722,No.81871954)the Beijing Natural Science Foundation(No.7212111)the Peking University Medicine Fund of Fostering Young Scholar’s Scientific&Technological Innovation supported by"the Fundamental Research Funds for the Central Universities"(BMU2018PY026)。
文摘Background:Cell competition is an important feature in pancreatic cancer(PC)progression,but the underlying mechanism remains elusive.This study aims to explore the role of exosomes derived from normal pancreatic ductal epithelial cells involved in PC progression.Methods:PC cells and pancreatic stellate cells(PSCs)were treated with exosomes isolated from pancreatic ductal epithelial cells.Cell proliferation was assessed by CCK8 assays.Cell migration and invasion were assessed by Transwell assays.PC and matched adjacent non-tumor tissue specimens were obtained from 46 patients pathologically diagnosed with PC at Peking University First Hospital from 2013 to 2017.Tissue miR-485-3p and p21-activated kinase-1(PAK1)expression was examined by real-time polymerase chain reaction(RT-PCR),and the relationship of the two was analyzed using Pearman’s product-moment correlation.The clinical significance of miR-485-3p was analyzed using the Chi-square test,Wilcoxon rank-sum test,and Fisher exact probability,respectively.The binding of miR-485-3p to PAK15’-untranslated region(5’-UTR)was examined by luciferase assay.PC cells were xenografted into nude mice as a PC metastasis model.Results:Exosomes from pancreatic ductal epithelial cells suppressed PC cell migration and invasion as well as the secretion and migration of PSCs.MiR-485-3p was enriched in the exosomes of pancreatic ductal epithelial cells but deficient in those of PC cells and PSCs,in accordance with the lower level in PSCs and PC cells than that in pancreatic ductal cells.And the mature miR-485-3p could be delivered into these cells by the exosomes secreted by normal pancreatic duct cells,to inhibit PC cell migration and invasion.Clinical data analysis showed that miR-485-3p was significantly decreased in PC tissues(P<0.05)and was negatively associated with lymphovascular invasion(P=0.044).As a direct target of miR-485-3p,PAK1 was found to exert an inhibitory effect on PC cells,and there was a significantly negative correlation between the expression levels of miR-485-3p and PAK1(r=-0.6525,P<0.0001)in PC tissues.Moreover,miR-485-3p could suppress PC metastasisin vivo by targeting p21-activated kinase-1.Conclusions:Exosomal miR-485-3p delivered by normal pancreatic ductal epithelial cells into PC cells inhibits PC metastasis by directly targeting PAK1.The restoration of miR-485-3p by exosomes or some other vehicle might be a novel approach for PC treatment.
基金National Natural Science Foundation of China(No. 81970344)
文摘Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI.However,the mechanisms underlying the regulatory effect of autophagy in AKI among patients undergoing cardiac surgeries are poorly understood.In this study,we aimed to test the hypothesis that exosomal microRNAs(miRNAs)regulate autophagy in tubular epithelial cells after AKI.Methods:Plasma exosomal RNA was extracted from young and elderly AKI patients undergoing cardiac surgery,and the miRNAs expression during the perioperative period were analyzed using next-generation sequencing.The screened miRNAs and their target genes were subjected to gene oncology function and Kyoto Encyclopedia of Genes and Genome enrichment analyses.Renal tubular epithelial cell line(HK-2 cells)was cultured and hypoxia/reoxygenation(H/R)model was established,which is an in vitro renal ischemia/reperfusion(I/R)model.We used Western blot analysis,cell viability assay,transfection,luciferase assay to investigate the mechanisms underlying the observed increases in the levels of renal I/R injury-mediated exosomal miRNAs and their roles in regulating HK-2 cells autophagy.Results:miR-590-3p was highly enriched in the plasma exosomes of young AKI patients after cardiac surgery.Increased levels of miR-590-3p led to the increases in the expression of autophagy marker proteins,including Beclin-1 and microtubule associated protein 1 light chain 3 beta(LC3II),and prolonged the autophagic response in HK-2 cells after H/R treatment.These effects were achieved mainly via increases in the exosomal miR-590-3p levels,and the tumor necrosis factor receptor-associated factor 6 protein was shown to play a key role in I/R injury-mediated autophagy induction.Conclusion:Exosomes released from HK-2 cells after renal I/R injury regulate autophagy by transferring miR-590-3p in a paracrine manner,which suggests that increasing the miR-590-3p levels in HK-2 cell-derived exosomes may increase autophagy and protect against kidney injury after renal I/R injury.
基金the National Natural Science Foundation of China(31371180)。
文摘Glioma-associated microglial cells,a key component of the tumor microenvironment,play an important role in glioma progression.In this study,the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were chosen.First,circadian gene expression in glioma cells co-cultured with either M1 or M2 microglia was assessed and the exosomes of M2-polarized and unpolarized BV-2 microglia were extracted.Subsequently,we labeled the exosomes with PKH67 and treated GL261 cells with them to investigate the exosome distribution.GL261 cell phenotypes and related protein expression were used to explore the role of M2 microglial exosomes in gliomas.Then a specific miR-7239-3p inhibitor was added to verify miR-7239-3p functions.Finally,the mouse subcutaneous tumorigenic model was used to verify the tumorigenic effect of M2 microglial exosomes in vivo.Our results showed that in gliomas co-cultured with M2 microglia,the expression of the BMAL1 protein was decreased(P<0.01),while the expression of the CLOCK protein was increased(P<0.05);opposite results were obtained in gliomas co-cultured with M1 microglia.After treatment with M2 microglial exosomes,the apoptosis of GL261 cells decreased(P<0.001),while the viability,proliferation,and migration of GL261 cells increased.Increased expression of N-cadherin and Vimentin,and decreased E-cadherin expression occurred upon treatment with M2 microglial exosomes.Addition of an miR-7239-3p inhibitor to M2 microglial exosomes reversed these results.In summary,we found that miR-7239-3p in the glioma microenvironment is recruited to glioma cells by exosomes and inhibits Bmal1 expression.M2 microglial exosomes promote the proliferation and migration of gliomas by regulating tumor-related protein expression and reducing apoptosis.
