Background:MiRNAs act as pivotal post-transcriptional gene mediators in the regulation of diverse biological processes,including proliferation,development and apoptosis.Our previous study has showed that miR-101-3p is...Background:MiRNAs act as pivotal post-transcriptional gene mediators in the regulation of diverse biological processes,including proliferation,development and apoptosis.Our previous study has showed that miR-101-3p is differentially expressed in dairy goat ovaries compared single with multiple litters.The objective of this research was to explore the potential function and molecular mechanism of miR-101-3p via its target STC1 in goat ovarian growth and development.Results:cDNA libraries were constructed using goat granulosa cells transfected with miR-101-3p mimics and negative control by RNA-sequencing.In total,142 differentially expressed unigenes(DEGs)were detected between two libraries,including 78 down-regulated and 64 up-regulated genes.GO and KEGG enrichment analysis showed the potential impacts of DEGs on ovarian development.STC1 was singled out from DEGs for further research owing to it regulates reproductive-related processes.In vitro,bioinformatics analysis and 3′-UTR assays confirmed that STC1 was a target of miR-101-3p.ELISA was performed to detect the estrogen(E2)and progesterone(P4)levels.CCK8,EdU and flow cytometry assays were performed to detect the proliferation and apoptosis of granulosa cells.Results showed that miR-101-3p regulated STAR,CYP19A1,CYP11A1 and 3β-HSD steroid hormone synthesis-associated genes by STC1 depletion,thus promoted E2 and P4 secretions.MiR-101-3p also affected the key protein PI3K,PTEN,AKT and mTOR in PI3K-AKT pathway by STC1,thereby suppressing proliferation and promoting apoptosis of granulosa cells.In vivo,the distribution and expression levels of miR-101-3p in mouse ovaries were determined through fluorescence in situ hybridisation(FISH).Immunohistochemistry results showed that STC1 expression was suppressed in mouse ovaries in miR-101-3p-agonist and siRNA-STC1 groups.Small and stunted ovarian fragments,decreased numbers of follicles at diverse stages were observed using Hematoxylin-eosin(HE)staining,thereby showing unusual ovarian development after miR-101-3p overexpression or STC1 depletion.Inhibition of miR-101-3p manifested opposite results.Conclusions:Taken together,our results demonstrated a regulatory mechanism of miR-101-3p via STC1 in goat granulosa cells,and offered the first in vivo example of miR-101-3p and STC1 functions required for ovarian development.展开更多
目的:观察斯钙素-1(STC-1)在喉癌患者和健康人群血清中的表达,探讨STC-1与喉癌患者临床病理特征的关系及对预后判断的指导意义。方法:应用Real time PCR和ELISA方法从基因水平和蛋白水平对喉癌患者及健康人群进行STC-1表达分析,并结合...目的:观察斯钙素-1(STC-1)在喉癌患者和健康人群血清中的表达,探讨STC-1与喉癌患者临床病理特征的关系及对预后判断的指导意义。方法:应用Real time PCR和ELISA方法从基因水平和蛋白水平对喉癌患者及健康人群进行STC-1表达分析,并结合患者病理指标、病理分型和临床随访资料进行回顾性分析。结果:基因水平和蛋白水平检测发现STC-1在喉癌患者血清中表达高于健康对照人群血清中的表达。STC-1表达与病理分化等喉癌血清学指标有一定的相关性,且与喉癌的大小、是否转移及临床分型等密切相关。结论:首次发现STC-1在喉癌血清中高表达,且其表达水平与喉癌的病理分期及不良预后密切相关,是喉癌潜在的诊断预警指标物。展开更多
基金supported by the National Natural Science Foundation of China(31601925)Shaanxi Science and Technology Innovation Project Plan(2020ZDLNY02–01,2020ZDLNY02–02,2018ZDCXL-NY-01-04,2018ZDCXL-NY-01-02 and 2017ZDXM-NY-081)Natural Science Foundation of Shaanxi Province(2020JQ-868)。
文摘Background:MiRNAs act as pivotal post-transcriptional gene mediators in the regulation of diverse biological processes,including proliferation,development and apoptosis.Our previous study has showed that miR-101-3p is differentially expressed in dairy goat ovaries compared single with multiple litters.The objective of this research was to explore the potential function and molecular mechanism of miR-101-3p via its target STC1 in goat ovarian growth and development.Results:cDNA libraries were constructed using goat granulosa cells transfected with miR-101-3p mimics and negative control by RNA-sequencing.In total,142 differentially expressed unigenes(DEGs)were detected between two libraries,including 78 down-regulated and 64 up-regulated genes.GO and KEGG enrichment analysis showed the potential impacts of DEGs on ovarian development.STC1 was singled out from DEGs for further research owing to it regulates reproductive-related processes.In vitro,bioinformatics analysis and 3′-UTR assays confirmed that STC1 was a target of miR-101-3p.ELISA was performed to detect the estrogen(E2)and progesterone(P4)levels.CCK8,EdU and flow cytometry assays were performed to detect the proliferation and apoptosis of granulosa cells.Results showed that miR-101-3p regulated STAR,CYP19A1,CYP11A1 and 3β-HSD steroid hormone synthesis-associated genes by STC1 depletion,thus promoted E2 and P4 secretions.MiR-101-3p also affected the key protein PI3K,PTEN,AKT and mTOR in PI3K-AKT pathway by STC1,thereby suppressing proliferation and promoting apoptosis of granulosa cells.In vivo,the distribution and expression levels of miR-101-3p in mouse ovaries were determined through fluorescence in situ hybridisation(FISH).Immunohistochemistry results showed that STC1 expression was suppressed in mouse ovaries in miR-101-3p-agonist and siRNA-STC1 groups.Small and stunted ovarian fragments,decreased numbers of follicles at diverse stages were observed using Hematoxylin-eosin(HE)staining,thereby showing unusual ovarian development after miR-101-3p overexpression or STC1 depletion.Inhibition of miR-101-3p manifested opposite results.Conclusions:Taken together,our results demonstrated a regulatory mechanism of miR-101-3p via STC1 in goat granulosa cells,and offered the first in vivo example of miR-101-3p and STC1 functions required for ovarian development.
文摘目的:观察斯钙素-1(STC-1)在喉癌患者和健康人群血清中的表达,探讨STC-1与喉癌患者临床病理特征的关系及对预后判断的指导意义。方法:应用Real time PCR和ELISA方法从基因水平和蛋白水平对喉癌患者及健康人群进行STC-1表达分析,并结合患者病理指标、病理分型和临床随访资料进行回顾性分析。结果:基因水平和蛋白水平检测发现STC-1在喉癌患者血清中表达高于健康对照人群血清中的表达。STC-1表达与病理分化等喉癌血清学指标有一定的相关性,且与喉癌的大小、是否转移及临床分型等密切相关。结论:首次发现STC-1在喉癌血清中高表达,且其表达水平与喉癌的病理分期及不良预后密切相关,是喉癌潜在的诊断预警指标物。
