Chamber maturation is a significant process in cardiac development. Disorders of this crucial process lead to a range of congenital heart defects. Foxc1a is a critical transcription factor reported to regulate the spe...Chamber maturation is a significant process in cardiac development. Disorders of this crucial process lead to a range of congenital heart defects. Foxc1a is a critical transcription factor reported to regulate the specification of cardiac progenitor cells. However, little is known about the role of Foxc1a in modulating chamber maturation. Previously, we reported that foxc1a-null zebrafish embryos exhibit disrupted heart structures and functions. In this study, we observe that ventricle structure and cardiomyocyte proliferation are abolished during chamber maturation in foxc1a-null zebrafish embryos. To observe the endogenous localization of Foxc1a in the hearts of living embryos, we insert eyfp at the foxc1a genomic locus using TALEN. Analysis of the knockin zebrafish show that foxc1a is widely expressed in ventricular cardiomyocytes during chamber development. Cardiac RNA sequencing analysis reveals the downregulated expression of the Hippo signaling effector wwtr1. Dual-luciferase and chromatin immunoprecipitation assays reveal that Foxc1a can bind directly to three sites in the wwtr1 promoter region. Furthermore, wwtr1m RNA overexpression is sufficient to reverse the ventricle defects during chamber maturation. Conditional overexpression of nkx2.5 also partially rescues the ventricular defects during chamber development. These findings demonstrate that wwtr1 and nkx2.5 are direct targets of Foxc1a during ventricular chamber maturation.展开更多
目的:探讨FOXC1蛋白表达与子宫颈癌及子宫内膜癌的临床病理特征间相关性。方法:采用免疫组化S-P法检测54例子宫颈癌及23例子宫内膜癌石蜡组织中FOXC1蛋白的表达。结果:FOXC1蛋白分别在29(53.7%)例子宫颈癌组织及20(87.0%)例子宫内膜癌...目的:探讨FOXC1蛋白表达与子宫颈癌及子宫内膜癌的临床病理特征间相关性。方法:采用免疫组化S-P法检测54例子宫颈癌及23例子宫内膜癌石蜡组织中FOXC1蛋白的表达。结果:FOXC1蛋白分别在29(53.7%)例子宫颈癌组织及20(87.0%)例子宫内膜癌组织中均有表达,且定位在细胞浆。FOXC1阳性表达与宫颈癌组织学分级相关(P<0.05),与宫颈癌的病理分型,FIGO(国际妇产科联盟International Federation of Gynecology and Obstetrics)临床分期,宫颈肌层浸润及盆腔淋巴结转移之间无明显相关性(P>0.05)。FOXC1蛋白表达与子宫内膜癌病理类型,组织学分级,FIGO手术病理分期,肌层浸润深度,宫颈受累情况及附件受累,盆腔淋巴结转移之间均无明显相关性(P>0.05)。结论:FOXC1蛋白可能参与了子宫颈癌的发生及发展过程,它可能与子宫内膜癌发生发展过程无关。展开更多
目的:研究小干扰RNA(small interfering R N A,s i R N A)沉默人胰腺癌Capan-2、PANC-1细胞中叉头框蛋白C1(fork head box C1,FOXC1)基因对胰腺癌细胞增殖能力的影响及作用机制.方法:以胰腺癌细胞、原发胰腺癌组织为研究对象,实时定量...目的:研究小干扰RNA(small interfering R N A,s i R N A)沉默人胰腺癌Capan-2、PANC-1细胞中叉头框蛋白C1(fork head box C1,FOXC1)基因对胰腺癌细胞增殖能力的影响及作用机制.方法:以胰腺癌细胞、原发胰腺癌组织为研究对象,实时定量逆转录聚合酶链反应(quantitative real-time reverse transcription polymerase chain reaction,q RT-PCR)检测FOXC1 m RNA在胰腺癌细胞及组织中的表达情况;将人胰腺癌细胞分为2组:FOXC1s i R N A组(实验组)、N C s i R N A组(阴性对照组),脂质体转染法将FOXC1 si RNA转染入胰腺癌细胞,q RT-PCR、Western blot技术检测胰腺癌细胞FOXC1 m RNA及蛋白表达变化,5-乙炔基-2'-脱氧尿苷(5-ethynyl-2'-deoxyuridine,Ed U)细胞增殖法检测各组胰腺癌细胞增殖情况;流式细胞技术(flow cytometry,FCM)检测各组胰腺癌细胞的周期分布,Western blot技术分析周期相关蛋白P21、P53、Cyclin D1蛋白表达情况.结果:q RT-PCR结果提示:相比正常胰腺上皮细胞和癌旁胰腺组织,FOXC1 m RNA在胰腺癌细胞及胰腺癌组织中表达较高(P<0.05);q RT-P C R及Westernblo t结果显示FOXC1 si RNA有效沉默了胰腺癌细胞F O X C1基因的转录和表达,E d U细胞增殖实验提示:沉默了胰腺癌细胞F O X C1基因表达后,胰腺癌细胞胞的增殖能力明显下降,较阴性对照组,差异有统计学意义(P<0.05);F C M结果显示:沉默了胰腺癌细胞FOXC1基因表达后胰腺癌细胞被阻滞在G0/G1期,较阴性对照组,差异有统计学意义(P<0.05);Western blot结果显示:较阴性对照组,P21、P53表达水平无明显变化,Cyclin D1表达水平下降,差异有统计学意义(P<0.05).结论:FOXC1 si RNA能够有效沉默人胰腺癌Capan-2、PANC-1细胞FOXC1基因的表达,抑制其增殖,将细胞周期阻滞于G0/G1期,提示FOXC1影响细胞增殖可能是通过调控细胞周期实现的,其机制可能是部分通过调控细胞周期蛋白Cyclin D1的表达而实现.展开更多
目的:探讨FOXC1基因过表达对口腔鳞癌细胞SCC-9增殖、凋亡及迁移能力的影响。方法:运用PCR技术对目的基因FOXC1进行扩增,进一步应用基因重组技术,将靶基因克隆入慢病毒载体,构建含有FOXC1基因的重组慢病毒载体。将构建载体转染至SCC-9细...目的:探讨FOXC1基因过表达对口腔鳞癌细胞SCC-9增殖、凋亡及迁移能力的影响。方法:运用PCR技术对目的基因FOXC1进行扩增,进一步应用基因重组技术,将靶基因克隆入慢病毒载体,构建含有FOXC1基因的重组慢病毒载体。将构建载体转染至SCC-9细胞,分别应用流式细胞术、R T-PCR以及Wester n blot检测SCC-9细胞中FOXC1 m R NA和蛋白表达水平,验证转染效率。最后通过Annex in V/7-AAD、CCK-8细胞活力检测、细胞周期检测及细胞划痕试验分别评估FOXC1基因过表达对SCC-9细胞增殖、凋亡及迁移能力的影响。结果:与空白对照组及空载体对照组相比,实验组(过表达FOXC1组)细胞FOXC1的m RNA及蛋白表达水平均较有所升高(P<0.05);实验组SCC-9细胞的增殖和迁移能力均有所减弱,凋亡细胞比例显著增加(P<0.05)。结论:过表达FOXC1基因能够在一定程度上抑制口腔鳞癌细胞SCC-9增殖和迁移能力,促进肿瘤细胞凋亡。展开更多
基金supported by the National Natural Science Foundation of China (31970769 and 31671518)。
文摘Chamber maturation is a significant process in cardiac development. Disorders of this crucial process lead to a range of congenital heart defects. Foxc1a is a critical transcription factor reported to regulate the specification of cardiac progenitor cells. However, little is known about the role of Foxc1a in modulating chamber maturation. Previously, we reported that foxc1a-null zebrafish embryos exhibit disrupted heart structures and functions. In this study, we observe that ventricle structure and cardiomyocyte proliferation are abolished during chamber maturation in foxc1a-null zebrafish embryos. To observe the endogenous localization of Foxc1a in the hearts of living embryos, we insert eyfp at the foxc1a genomic locus using TALEN. Analysis of the knockin zebrafish show that foxc1a is widely expressed in ventricular cardiomyocytes during chamber development. Cardiac RNA sequencing analysis reveals the downregulated expression of the Hippo signaling effector wwtr1. Dual-luciferase and chromatin immunoprecipitation assays reveal that Foxc1a can bind directly to three sites in the wwtr1 promoter region. Furthermore, wwtr1m RNA overexpression is sufficient to reverse the ventricle defects during chamber maturation. Conditional overexpression of nkx2.5 also partially rescues the ventricular defects during chamber development. These findings demonstrate that wwtr1 and nkx2.5 are direct targets of Foxc1a during ventricular chamber maturation.
文摘目的:探讨FOXC1蛋白表达与子宫颈癌及子宫内膜癌的临床病理特征间相关性。方法:采用免疫组化S-P法检测54例子宫颈癌及23例子宫内膜癌石蜡组织中FOXC1蛋白的表达。结果:FOXC1蛋白分别在29(53.7%)例子宫颈癌组织及20(87.0%)例子宫内膜癌组织中均有表达,且定位在细胞浆。FOXC1阳性表达与宫颈癌组织学分级相关(P<0.05),与宫颈癌的病理分型,FIGO(国际妇产科联盟International Federation of Gynecology and Obstetrics)临床分期,宫颈肌层浸润及盆腔淋巴结转移之间无明显相关性(P>0.05)。FOXC1蛋白表达与子宫内膜癌病理类型,组织学分级,FIGO手术病理分期,肌层浸润深度,宫颈受累情况及附件受累,盆腔淋巴结转移之间均无明显相关性(P>0.05)。结论:FOXC1蛋白可能参与了子宫颈癌的发生及发展过程,它可能与子宫内膜癌发生发展过程无关。
文摘目的:研究小干扰RNA(small interfering R N A,s i R N A)沉默人胰腺癌Capan-2、PANC-1细胞中叉头框蛋白C1(fork head box C1,FOXC1)基因对胰腺癌细胞增殖能力的影响及作用机制.方法:以胰腺癌细胞、原发胰腺癌组织为研究对象,实时定量逆转录聚合酶链反应(quantitative real-time reverse transcription polymerase chain reaction,q RT-PCR)检测FOXC1 m RNA在胰腺癌细胞及组织中的表达情况;将人胰腺癌细胞分为2组:FOXC1s i R N A组(实验组)、N C s i R N A组(阴性对照组),脂质体转染法将FOXC1 si RNA转染入胰腺癌细胞,q RT-PCR、Western blot技术检测胰腺癌细胞FOXC1 m RNA及蛋白表达变化,5-乙炔基-2'-脱氧尿苷(5-ethynyl-2'-deoxyuridine,Ed U)细胞增殖法检测各组胰腺癌细胞增殖情况;流式细胞技术(flow cytometry,FCM)检测各组胰腺癌细胞的周期分布,Western blot技术分析周期相关蛋白P21、P53、Cyclin D1蛋白表达情况.结果:q RT-PCR结果提示:相比正常胰腺上皮细胞和癌旁胰腺组织,FOXC1 m RNA在胰腺癌细胞及胰腺癌组织中表达较高(P<0.05);q RT-P C R及Westernblo t结果显示FOXC1 si RNA有效沉默了胰腺癌细胞F O X C1基因的转录和表达,E d U细胞增殖实验提示:沉默了胰腺癌细胞F O X C1基因表达后,胰腺癌细胞胞的增殖能力明显下降,较阴性对照组,差异有统计学意义(P<0.05);F C M结果显示:沉默了胰腺癌细胞FOXC1基因表达后胰腺癌细胞被阻滞在G0/G1期,较阴性对照组,差异有统计学意义(P<0.05);Western blot结果显示:较阴性对照组,P21、P53表达水平无明显变化,Cyclin D1表达水平下降,差异有统计学意义(P<0.05).结论:FOXC1 si RNA能够有效沉默人胰腺癌Capan-2、PANC-1细胞FOXC1基因的表达,抑制其增殖,将细胞周期阻滞于G0/G1期,提示FOXC1影响细胞增殖可能是通过调控细胞周期实现的,其机制可能是部分通过调控细胞周期蛋白Cyclin D1的表达而实现.
文摘目的:探讨FOXC1基因过表达对口腔鳞癌细胞SCC-9增殖、凋亡及迁移能力的影响。方法:运用PCR技术对目的基因FOXC1进行扩增,进一步应用基因重组技术,将靶基因克隆入慢病毒载体,构建含有FOXC1基因的重组慢病毒载体。将构建载体转染至SCC-9细胞,分别应用流式细胞术、R T-PCR以及Wester n blot检测SCC-9细胞中FOXC1 m R NA和蛋白表达水平,验证转染效率。最后通过Annex in V/7-AAD、CCK-8细胞活力检测、细胞周期检测及细胞划痕试验分别评估FOXC1基因过表达对SCC-9细胞增殖、凋亡及迁移能力的影响。结果:与空白对照组及空载体对照组相比,实验组(过表达FOXC1组)细胞FOXC1的m RNA及蛋白表达水平均较有所升高(P<0.05);实验组SCC-9细胞的增殖和迁移能力均有所减弱,凋亡细胞比例显著增加(P<0.05)。结论:过表达FOXC1基因能够在一定程度上抑制口腔鳞癌细胞SCC-9增殖和迁移能力,促进肿瘤细胞凋亡。
