Mint is a newly identified molecule that mediates signal transduction and modulates chromatin repression. Mint family members contain a highly conserved C-terminus SPOC domain (SpenParalog and OrthologsC-terminal doma...Mint is a newly identified molecule that mediates signal transduction and modulates chromatin repression. Mint family members contain a highly conserved C-terminus SPOC domain (SpenParalog and OrthologsC-terminal domain) commonly associated with proliferation and related diseases (for example: cancer) due to its role in cell differentiation and apoptosis. In this study, we addressed the SPOC function using a tetracycline-inducible system to express the target domain in Ain V15 embryonic ES cells and bone marrow stem cells from SPOC transenic mice. In vitro differentiation of Ain V15 ES cells as a model of early hematopoietic development, we found expression of SPOC domain induces hematopoietic differentiation via up-regulation of transcription factors Bmp4 and Smad5, which induce the expression of hematopoietic factors Eklf1 and hematopoietic proliferation associated factor Gata2, the SPOC domain also plays the regulation function in the differentiation of hematopoitic progenitor by colony forming Unit (CFU) assays. Further, we determined SPOC expression enhances erythrocyte and granulocyte maturationusing bone marrow cells derived from tiSPOC chimeric mice. Finally, we identified that overexpression of full length Mint in ES cells drive Smad5 and Bmp4 up-regulation under culture conditions, and up-regulation of endogenous Mint when induceshematopoitic differentiation of EML, M1 and WT18 cells. In summary, our study reveals the conserved SPOC domain of Mint protein induces differentiation both in the stages of embryonic stem cells and hematopoietic progenitor cells.展开更多
目的探讨骨形态形成蛋白4(bone morphogenetic protein 4,BMP4)和Smad4蛋白(drosophila mothers against decapentaplegic protein 4)在C57BL/6小鼠内耳前庭发育不同阶段的表达。方法选择从胚胎第10天(E10)到胚胎第20天(E20)每天的孕鼠...目的探讨骨形态形成蛋白4(bone morphogenetic protein 4,BMP4)和Smad4蛋白(drosophila mothers against decapentaplegic protein 4)在C57BL/6小鼠内耳前庭发育不同阶段的表达。方法选择从胚胎第10天(E10)到胚胎第20天(E20)每天的孕鼠各两只(共22只),取E10～E17的胚胎头、E18～E20的胚胎内耳,通过冰冻连续切片、免疫组化或免疫荧光染色,观察小鼠胚胎内耳前庭发育过程中BMP4/Smad4的表达情况。结果 BMP4从E10开始,在间充质浓聚前就有表达;间充质一开始浓聚,BMP4在其中就有较丰富的表达;早期主要表达在前庭囊及前庭囊周围的间充质,在胚胎发育的后期,其在软骨囊及前庭上皮里有广泛阳性表达;Smad4在小鼠内耳前庭的表达情况与BMP4不一致,E10到E12的小鼠内耳Smad4表达阴性,E15时各个壶腹嵴、囊斑里才有较明显的Smad4表达,而内耳周围软骨囊到E16时才有明显的Smad4表达。结论 BMP4与小鼠内耳前庭形态形成及毛细胞分化发育、功能形成有关;Smad4与BMP4的表达时空特征不同,Smad4可能主要参与了小鼠内耳前庭发育后期的形态塑形和功能发育。展开更多
The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(G...The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(GC)and the underlying mechanisms.The differentially expressed miRNAs were firstly identified in serum exosomes of GC patients and healthy individuals using next-generation sequencing and bioinformatics.Next,the expression of serum exosomal miR-4256 was analyzed in GC cells and GC tissues,and the role of miR-4256 in GC was investigated by in vitro and in vivo experiments.Then,the effect of miR-4256 on its downstream target genes HDAC5/p16^(INK4a) was studied in GC cells,and the underlying mechanisms were evaluated using dual luciferase reporter assay and Chromatin Immunoprecipitation(ChIP).Additionally,the role of the miR-4256/HDAC5/p16^(INK4a) axis in GC was studied using in vitro and in vivo experiments.Finally,the upstream regulators SMAD2/p300 that regulate miR-4256 expression and their role in GC were explored using in vitro experiments.miR-4256 was the most significantly upregulated miRNA and was overexpressed in GC cell lines and GC tissues;in vitro and in vivo results showed that miR-4256 promoted GC growth and progression.Mechanistically,miR-4256 enhanced HDAC5 expression by targeting the promoter of the HDAC5 gene in GC cells,and then restrained the expression of p16^(INK4a) through the epigenetic modulation of HDAC5 at the p16INK4a promoter.Furthermore,miR-4256 overexpression was positively regulated by the SMAD2/p300 complex in GC cells.Our data indicate that miR-4256 functions as an oncogene in GC via the SMAD2/miR-4256/HDAC5/p16^(INK4a) axis,which participates in GC progression and provides novel therapeutic and prognostic biomarkers for GC.展开更多
试验旨在探索骨形态发生蛋白4(Bone morphogenetic protein 4,BMP4)和SMAD家族蛋白4(SMAD family member 4,SMAD4)基因在公绵羊繁殖中的作用,以便深入研究公绵羊繁殖机制。采用荧光定量PCR技术检测BMP4和SMAD4基因在高繁殖力的小尾寒羊...试验旨在探索骨形态发生蛋白4(Bone morphogenetic protein 4,BMP4)和SMAD家族蛋白4(SMAD family member 4,SMAD4)基因在公绵羊繁殖中的作用,以便深入研究公绵羊繁殖机制。采用荧光定量PCR技术检测BMP4和SMAD4基因在高繁殖力的小尾寒羊公羊(n=3)和低繁殖力苏尼特公羊(n=3)大脑、小脑、下丘脑、垂体、输精管、肾上腺、睾丸和附睾8种组织中的表达。结果表明:BMP4和SMAD4基因在小尾寒羊公羊和苏尼特羊公羊各种组织中均有表达。其中BMP4基因在睾丸中高表达,在输精管、肾上腺中中等表达,在其他组织中均呈痕量表达;SMAD4基因在睾丸、下丘脑、小脑、大脑中高表达,在其他组织中中等表达。BMP4基因在低繁殖力苏尼特羊公羊睾丸中表达量显著高于高繁殖力小尾寒羊公羊(P<0.05);SMAD4基因在低繁殖力苏尼特公羊8种组织中表达量均显著高于高繁殖力小尾寒羊公羊(P<0.05)。说明BMP4和SMAD4基因在公绵羊繁殖中起到一定程度的负调控作用,这为公羊高繁殖性状选育提供了参考依据。展开更多
Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a nov...Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.展开更多
文摘Mint is a newly identified molecule that mediates signal transduction and modulates chromatin repression. Mint family members contain a highly conserved C-terminus SPOC domain (SpenParalog and OrthologsC-terminal domain) commonly associated with proliferation and related diseases (for example: cancer) due to its role in cell differentiation and apoptosis. In this study, we addressed the SPOC function using a tetracycline-inducible system to express the target domain in Ain V15 embryonic ES cells and bone marrow stem cells from SPOC transenic mice. In vitro differentiation of Ain V15 ES cells as a model of early hematopoietic development, we found expression of SPOC domain induces hematopoietic differentiation via up-regulation of transcription factors Bmp4 and Smad5, which induce the expression of hematopoietic factors Eklf1 and hematopoietic proliferation associated factor Gata2, the SPOC domain also plays the regulation function in the differentiation of hematopoitic progenitor by colony forming Unit (CFU) assays. Further, we determined SPOC expression enhances erythrocyte and granulocyte maturationusing bone marrow cells derived from tiSPOC chimeric mice. Finally, we identified that overexpression of full length Mint in ES cells drive Smad5 and Bmp4 up-regulation under culture conditions, and up-regulation of endogenous Mint when induceshematopoitic differentiation of EML, M1 and WT18 cells. In summary, our study reveals the conserved SPOC domain of Mint protein induces differentiation both in the stages of embryonic stem cells and hematopoietic progenitor cells.
文摘目的探讨骨形态形成蛋白4(bone morphogenetic protein 4,BMP4)和Smad4蛋白(drosophila mothers against decapentaplegic protein 4)在C57BL/6小鼠内耳前庭发育不同阶段的表达。方法选择从胚胎第10天(E10)到胚胎第20天(E20)每天的孕鼠各两只(共22只),取E10～E17的胚胎头、E18～E20的胚胎内耳,通过冰冻连续切片、免疫组化或免疫荧光染色,观察小鼠胚胎内耳前庭发育过程中BMP4/Smad4的表达情况。结果 BMP4从E10开始,在间充质浓聚前就有表达;间充质一开始浓聚,BMP4在其中就有较丰富的表达;早期主要表达在前庭囊及前庭囊周围的间充质,在胚胎发育的后期,其在软骨囊及前庭上皮里有广泛阳性表达;Smad4在小鼠内耳前庭的表达情况与BMP4不一致,E10到E12的小鼠内耳Smad4表达阴性,E15时各个壶腹嵴、囊斑里才有较明显的Smad4表达,而内耳周围软骨囊到E16时才有明显的Smad4表达。结论 BMP4与小鼠内耳前庭形态形成及毛细胞分化发育、功能形成有关;Smad4与BMP4的表达时空特征不同,Smad4可能主要参与了小鼠内耳前庭发育后期的形态塑形和功能发育。
基金The studies involving human participants were approved by The First Affiliated Hospital of Jinan University Ethics Committee(KY-2021-095)The participants provided their written informed consent to participate in this study+1 种基金Animalinvolved experimental protocols were compliance with guidelines and licensesapproved by the Laboratory Animal Center of Jinan University(20220225-65).
文摘The dysregulation of exosomal microRNAs(miRNAs)plays a crucial role in the development and progression of cancer.This study investigated the role of a newly identified serum exosomal miRNA miR-4256 in gastric cancer(GC)and the underlying mechanisms.The differentially expressed miRNAs were firstly identified in serum exosomes of GC patients and healthy individuals using next-generation sequencing and bioinformatics.Next,the expression of serum exosomal miR-4256 was analyzed in GC cells and GC tissues,and the role of miR-4256 in GC was investigated by in vitro and in vivo experiments.Then,the effect of miR-4256 on its downstream target genes HDAC5/p16^(INK4a) was studied in GC cells,and the underlying mechanisms were evaluated using dual luciferase reporter assay and Chromatin Immunoprecipitation(ChIP).Additionally,the role of the miR-4256/HDAC5/p16^(INK4a) axis in GC was studied using in vitro and in vivo experiments.Finally,the upstream regulators SMAD2/p300 that regulate miR-4256 expression and their role in GC were explored using in vitro experiments.miR-4256 was the most significantly upregulated miRNA and was overexpressed in GC cell lines and GC tissues;in vitro and in vivo results showed that miR-4256 promoted GC growth and progression.Mechanistically,miR-4256 enhanced HDAC5 expression by targeting the promoter of the HDAC5 gene in GC cells,and then restrained the expression of p16^(INK4a) through the epigenetic modulation of HDAC5 at the p16INK4a promoter.Furthermore,miR-4256 overexpression was positively regulated by the SMAD2/p300 complex in GC cells.Our data indicate that miR-4256 functions as an oncogene in GC via the SMAD2/miR-4256/HDAC5/p16^(INK4a) axis,which participates in GC progression and provides novel therapeutic and prognostic biomarkers for GC.
文摘试验旨在探索骨形态发生蛋白4(Bone morphogenetic protein 4,BMP4)和SMAD家族蛋白4(SMAD family member 4,SMAD4)基因在公绵羊繁殖中的作用,以便深入研究公绵羊繁殖机制。采用荧光定量PCR技术检测BMP4和SMAD4基因在高繁殖力的小尾寒羊公羊(n=3)和低繁殖力苏尼特公羊(n=3)大脑、小脑、下丘脑、垂体、输精管、肾上腺、睾丸和附睾8种组织中的表达。结果表明:BMP4和SMAD4基因在小尾寒羊公羊和苏尼特羊公羊各种组织中均有表达。其中BMP4基因在睾丸中高表达,在输精管、肾上腺中中等表达,在其他组织中均呈痕量表达;SMAD4基因在睾丸、下丘脑、小脑、大脑中高表达,在其他组织中中等表达。BMP4基因在低繁殖力苏尼特羊公羊睾丸中表达量显著高于高繁殖力小尾寒羊公羊(P<0.05);SMAD4基因在低繁殖力苏尼特公羊8种组织中表达量均显著高于高繁殖力小尾寒羊公羊(P<0.05)。说明BMP4和SMAD4基因在公绵羊繁殖中起到一定程度的负调控作用,这为公羊高繁殖性状选育提供了参考依据。
基金supported by grants from the National Natural Science Foundation (31872979, 31572366)the National Key Research and Development Program of China (2017YFD0502002)the National Basic Research Programs of China (2015CB943102)。
文摘Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.
