Background The proliferation of porcine ovarian granulosa cells(GCs)is essential to follicular development and the ubiquitin–proteasome system is necessary for maintaining cell cycle homeostasis.Previous studies foun...Background The proliferation of porcine ovarian granulosa cells(GCs)is essential to follicular development and the ubiquitin–proteasome system is necessary for maintaining cell cycle homeostasis.Previous studies found that the deubiquitinase ubiquitin carboxyl-terminal hydrolase 1(UCHL1)regulates female reproduction,especially in ovarian development.However,the mechanism by which UCHL1 regulates porcine GC proliferation remains unclear.Results UCHL1 overexpression promoted GC proliferation,and knockdown had the opposite effect.UCHL1 is directly bound to cyclin B1(CCNB1),prolonging the half-life of CCNB1 and inhibiting its degradation,thereby promoting GC proliferation.What's more,a flavonoid compound-isovitexin improved the enzyme activity of UCHL1 and promoted the proliferation of porcine GCs.Conclusions UCHL1 promoted the proliferation of porcine GCs by stabilizing CCNB1,and isovitexin enhanced the enzyme activity of UCHL1.These findings reveal the role of UCHL1 and the potential of isovitexin in regulating proliferation and provide insights into identifying molecular markers and nutrients that affect follicle development.展开更多
目的探讨microRNA-144细胞周期蛋白B1(miR-144CCNB1)信号调节通路在肝癌增殖、迁移和侵袭作用及其相关机制。方法以2011年1月~2016年1月我院收治的100例肝癌患者的肝癌组织和癌旁组织为研究标本,采用荧光定量PCR(qRT-PCR)法检测肝癌及...目的探讨microRNA-144细胞周期蛋白B1(miR-144CCNB1)信号调节通路在肝癌增殖、迁移和侵袭作用及其相关机制。方法以2011年1月~2016年1月我院收治的100例肝癌患者的肝癌组织和癌旁组织为研究标本,采用荧光定量PCR(qRT-PCR)法检测肝癌及其癌旁组织的miR-144表达和CCNB1蛋白表达情况,采用RNA干扰技术沉默CCNB1基因,并将靶向CCNB1的siRNA转染至肝癌细胞系HepG2和QGY-7703中,检测其对肝癌细胞生长、迁移和侵袭的生物学行为的影响。结果肝癌组织中miR-144表达较癌旁组织明显下调,CCNB1蛋白在肝癌组织的表达水平较癌旁组织明显上调,miR-144高表达者预后无瘤生存期较其低表达者明显长,而CCNB1蛋白低表达者无瘤生存期明显长于其高表达者,差异有统计学意义(P<0.05);与未处理者和阴性对照模拟物相较,转染CCNB1 siR肝癌细胞CCNB1蛋白表达水平明显下降;与未处理和阴性对照模拟物相较,转染CCBN1 siR HepG2细胞凋亡百分明显升高,转染CCNB1 siR HepG2细胞增殖能力明显下降,转染CCNB1siR HepG2细胞迁移能力明显降低;与阴性对照模拟物和未处理细胞相较,转染CCNB1-siR QGY-7703细胞克隆团数(成瘤能力)明显减少。结论 miR-144CCNB1信号调节通路在肝癌增殖、迁移和侵袭中发挥重要作用,其可能机制为miR-144负向调控CCNB1表达,从而影响肝癌细胞增殖、凋亡、迁移等生物学行为,为肝癌治疗的新靶点提供理论依据。展开更多
基金funded by National Key R&D Program of China(NO.2022YFD1300303)National Natural Science Foundation of China(32272849)。
文摘Background The proliferation of porcine ovarian granulosa cells(GCs)is essential to follicular development and the ubiquitin–proteasome system is necessary for maintaining cell cycle homeostasis.Previous studies found that the deubiquitinase ubiquitin carboxyl-terminal hydrolase 1(UCHL1)regulates female reproduction,especially in ovarian development.However,the mechanism by which UCHL1 regulates porcine GC proliferation remains unclear.Results UCHL1 overexpression promoted GC proliferation,and knockdown had the opposite effect.UCHL1 is directly bound to cyclin B1(CCNB1),prolonging the half-life of CCNB1 and inhibiting its degradation,thereby promoting GC proliferation.What's more,a flavonoid compound-isovitexin improved the enzyme activity of UCHL1 and promoted the proliferation of porcine GCs.Conclusions UCHL1 promoted the proliferation of porcine GCs by stabilizing CCNB1,and isovitexin enhanced the enzyme activity of UCHL1.These findings reveal the role of UCHL1 and the potential of isovitexin in regulating proliferation and provide insights into identifying molecular markers and nutrients that affect follicle development.
文摘目的探讨microRNA-144细胞周期蛋白B1(miR-144CCNB1)信号调节通路在肝癌增殖、迁移和侵袭作用及其相关机制。方法以2011年1月~2016年1月我院收治的100例肝癌患者的肝癌组织和癌旁组织为研究标本,采用荧光定量PCR(qRT-PCR)法检测肝癌及其癌旁组织的miR-144表达和CCNB1蛋白表达情况,采用RNA干扰技术沉默CCNB1基因,并将靶向CCNB1的siRNA转染至肝癌细胞系HepG2和QGY-7703中,检测其对肝癌细胞生长、迁移和侵袭的生物学行为的影响。结果肝癌组织中miR-144表达较癌旁组织明显下调,CCNB1蛋白在肝癌组织的表达水平较癌旁组织明显上调,miR-144高表达者预后无瘤生存期较其低表达者明显长,而CCNB1蛋白低表达者无瘤生存期明显长于其高表达者,差异有统计学意义(P<0.05);与未处理者和阴性对照模拟物相较,转染CCNB1 siR肝癌细胞CCNB1蛋白表达水平明显下降;与未处理和阴性对照模拟物相较,转染CCBN1 siR HepG2细胞凋亡百分明显升高,转染CCNB1 siR HepG2细胞增殖能力明显下降,转染CCNB1siR HepG2细胞迁移能力明显降低;与阴性对照模拟物和未处理细胞相较,转染CCNB1-siR QGY-7703细胞克隆团数(成瘤能力)明显减少。结论 miR-144CCNB1信号调节通路在肝癌增殖、迁移和侵袭中发挥重要作用,其可能机制为miR-144负向调控CCNB1表达,从而影响肝癌细胞增殖、凋亡、迁移等生物学行为,为肝癌治疗的新靶点提供理论依据。
