BACKGROUND Pancreatic cancer(PC)is one of the deadliest cancers worldwide.PC metastasis involves a complex set of events,including epithelial-mesenchymal transition(EMT),that increase tumor cell invasiveness.Recent ev...BACKGROUND Pancreatic cancer(PC)is one of the deadliest cancers worldwide.PC metastasis involves a complex set of events,including epithelial-mesenchymal transition(EMT),that increase tumor cell invasiveness.Recent evidence has shown that hypoxia is a major EMT regulator in pancreatic cancer cells and facilitates metastasis;however,the mechanisms remain elusive.AIM To investigate the role of miR-301a in hypoxia-induced EMT in PC cells.METHODS Real-time PCR and Western blot analysis were used to detect the expression of miR-301a and EMT markers in PDAC cells cultured in hypoxic and normoxic conditions.Western blot analysis was used to detect the expression of EMT markers in PDAC cells with miR-301a overexpression.Wound healing assay and Transwell assay were used to detect the migration capabilities of PDAC cells with miR-301a overexpression and knockout.Luciferase assay was used to detect the miR-301a promoter and the 3’untranslated region activity of TP63.Orthotopic PC mouse models were used to study the role of miR-301a in metastasis of PDAC cells in vivo.In situ hybridization assay was used to detect the expression of miR-301a in PDAC patient samples(adjacent paratumor and paired tumor tissues).RESULTS Hypoxic environment could directly promote the EMT of PC cells.The expression level of miR-301a was increased in a HIF2αdependent manner in hypoxia-cultured CFPAC-1 and BxPC-3 cells.Overexpression of miR-301a enhanced the hypoxia-induced EMT of PC cells,while knocking out miR-301a result in the suppression of hypoxia-induced EMT.TP63 was a direct target of miR-301a and involved in the metastatic process of PC cells.Furthermore,miR-301a upregulation facilitated PDAC distant metastasis and lymph node metastasis in vivo.Additionally,miR-301a overexpression was indicative of aggressive clinicopathological behaviors and poor prognosis.CONCLUSION The newly identified HIF-2α-miR301a-TP63 signaling pathway may play a crucial role in hypoxia-induced EMT in PDAC cells.展开更多
BACKGROUND Cardiovascular disease is the leading cause of death worldwide.Tissue repair after pathological injury in the heart remains a major challenge due to the limited regenerative ability of cardiomyocytes in adu...BACKGROUND Cardiovascular disease is the leading cause of death worldwide.Tissue repair after pathological injury in the heart remains a major challenge due to the limited regenerative ability of cardiomyocytes in adults.Stem cell-derived cardiomyocytes provide a promising source for the cell transplantation-based treatment of injured hearts.AIM To explore the function and mechanisms of miR-301a in regulating cardiomyocyte differentiation of mouse embryonic stem(mES)cells,and provide experimental evidence for applying miR-301a to the cardiomyocyte differentiation induction from stem cells.METHODS mES cells with or without overexpression of miR-301a were applied for all functional assays.The hanging drop technique was applied to form embryoid bodies from mES cells.Cardiac markers including GATA-4,TBX5,MEF2C,andα-actinin were used to determine cardiomyocyte differentiation from mES cells.RESULTS High expression of miR-301a was detected in the heart from late embryonic to neonatal mice.Overexpression of miR-301a in mES cells significantly induced the expression of cardiac transcription factors,thereby promoting cardiomyocyte differentiation and beating cardiomyocyte clone formation.PTEN is a target gene of miR-301a in cardiomyocytes.PTEN-regulated PI3K-AKT-mTOR-Stat3 signaling showed involvement in regulating miR-301a-promoted cardiomyocyte differentiation from mES cells.CONCLUSION MiR-301a is capable of promoting embryonic stem cell differentiation to cardiomyocytes.展开更多
基金Supported by National Natural Science Foundation of China,No.81372640.
文摘BACKGROUND Pancreatic cancer(PC)is one of the deadliest cancers worldwide.PC metastasis involves a complex set of events,including epithelial-mesenchymal transition(EMT),that increase tumor cell invasiveness.Recent evidence has shown that hypoxia is a major EMT regulator in pancreatic cancer cells and facilitates metastasis;however,the mechanisms remain elusive.AIM To investigate the role of miR-301a in hypoxia-induced EMT in PC cells.METHODS Real-time PCR and Western blot analysis were used to detect the expression of miR-301a and EMT markers in PDAC cells cultured in hypoxic and normoxic conditions.Western blot analysis was used to detect the expression of EMT markers in PDAC cells with miR-301a overexpression.Wound healing assay and Transwell assay were used to detect the migration capabilities of PDAC cells with miR-301a overexpression and knockout.Luciferase assay was used to detect the miR-301a promoter and the 3’untranslated region activity of TP63.Orthotopic PC mouse models were used to study the role of miR-301a in metastasis of PDAC cells in vivo.In situ hybridization assay was used to detect the expression of miR-301a in PDAC patient samples(adjacent paratumor and paired tumor tissues).RESULTS Hypoxic environment could directly promote the EMT of PC cells.The expression level of miR-301a was increased in a HIF2αdependent manner in hypoxia-cultured CFPAC-1 and BxPC-3 cells.Overexpression of miR-301a enhanced the hypoxia-induced EMT of PC cells,while knocking out miR-301a result in the suppression of hypoxia-induced EMT.TP63 was a direct target of miR-301a and involved in the metastatic process of PC cells.Furthermore,miR-301a upregulation facilitated PDAC distant metastasis and lymph node metastasis in vivo.Additionally,miR-301a overexpression was indicative of aggressive clinicopathological behaviors and poor prognosis.CONCLUSION The newly identified HIF-2α-miR301a-TP63 signaling pathway may play a crucial role in hypoxia-induced EMT in PDAC cells.
基金Supported by the National Natural Science Foundation of China,No.81800243the Science and Technology Commission of Shanghai Municipality,No.18411965900the Fundamental Research Funds for the Central Universities,No.22120180125
文摘BACKGROUND Cardiovascular disease is the leading cause of death worldwide.Tissue repair after pathological injury in the heart remains a major challenge due to the limited regenerative ability of cardiomyocytes in adults.Stem cell-derived cardiomyocytes provide a promising source for the cell transplantation-based treatment of injured hearts.AIM To explore the function and mechanisms of miR-301a in regulating cardiomyocyte differentiation of mouse embryonic stem(mES)cells,and provide experimental evidence for applying miR-301a to the cardiomyocyte differentiation induction from stem cells.METHODS mES cells with or without overexpression of miR-301a were applied for all functional assays.The hanging drop technique was applied to form embryoid bodies from mES cells.Cardiac markers including GATA-4,TBX5,MEF2C,andα-actinin were used to determine cardiomyocyte differentiation from mES cells.RESULTS High expression of miR-301a was detected in the heart from late embryonic to neonatal mice.Overexpression of miR-301a in mES cells significantly induced the expression of cardiac transcription factors,thereby promoting cardiomyocyte differentiation and beating cardiomyocyte clone formation.PTEN is a target gene of miR-301a in cardiomyocytes.PTEN-regulated PI3K-AKT-mTOR-Stat3 signaling showed involvement in regulating miR-301a-promoted cardiomyocyte differentiation from mES cells.CONCLUSION MiR-301a is capable of promoting embryonic stem cell differentiation to cardiomyocytes.
