目的:分析chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)基因在结直肠癌组织中的表达情况及CHD1L基因表达对结直肠癌细胞侵袭和转移能力的影响.方法:收集结直肠癌患者癌和癌旁组织,通过荧光定量PCR、免疫组织...目的:分析chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)基因在结直肠癌组织中的表达情况及CHD1L基因表达对结直肠癌细胞侵袭和转移能力的影响.方法:收集结直肠癌患者癌和癌旁组织,通过荧光定量PCR、免疫组织化学及Western blot检测CHD1L的表达情况,运用Transwell实验、划痕实验研究CHD1L对结直肠癌细胞侵袭和转移能力的影响.结果:CHD1L mRNA和蛋白在结直肠癌组织中表达水平显著高于与之对应的癌旁组织,差异有统计学意义;此外,结肠癌细胞株中CHD1L的表达普遍呈现高表达.结肠癌细胞LOVO和DLD-1中转染shRNA-CHD1L后,可以明显降低CHD1L的mRNA和蛋白的表达,同时明显抑制结肠癌细胞的侵袭和迁移能力.体内实验进一步发现降低SW620细胞中CHD1L的表达可以明显抑制肿瘤的肝、肺转移.结论:CHD1L在结直肠癌组织和细胞中呈现高表达,CHD1L在结直肠癌的侵袭和转移过程中发挥重要作用.CHD1L有可能成为治疗结直肠癌的新的靶点.展开更多
目的:检测chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)在结直肠组织中的表达情况及探讨CHD1L基因对结直肠癌(colorectal cancer,CRC)细胞增殖和凋亡的影响.方法:通过荧光定量PCR、免疫组织化学检测80例CRC患...目的:检测chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)在结直肠组织中的表达情况及探讨CHD1L基因对结直肠癌(colorectal cancer,CRC)细胞增殖和凋亡的影响.方法:通过荧光定量PCR、免疫组织化学检测80例CRC患者癌组织和对应癌旁组织中CHD1L基因的表达情况;采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)、克隆形成实验及动物实验等观察CHD1L基因在CRC细胞增殖中的作用;利用流式细胞技术检测细胞凋亡.结果:76.25%(61/80)的CRC癌组织CHD1L基因的表达明显高于对应的癌旁组织;转染s h R N A-C H D1L至C R C细胞可以明显降低C H D1L基因的m R N A和蛋白的表达,同时M T T实验和克隆形成实验发现干扰C H D1L组细胞的增殖明显低于对照组.另外,流式细胞仪检测发现干扰CHD1L组较对照组细胞凋亡指数明显增加.裸鼠成瘤实验发现稳定低表达CHD1L组肿瘤体积和重量明显小于对照组.结论:CHD1L在结直肠组织呈现高表达,降低结直肠细胞中CHD1L的表达可明显抑制细胞增殖及肿瘤生长.CHD1L可能成为治疗CRC的新靶点.展开更多
Chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)是一种新发现的位于染色体1q21致癌基因,主要通过细胞增殖、G1/S期的转变、抑制凋亡、染色质不稳定等机制致癌,并被认为是影响癌症的进程、预后和生存率的独立标...Chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)是一种新发现的位于染色体1q21致癌基因,主要通过细胞增殖、G1/S期的转变、抑制凋亡、染色质不稳定等机制致癌,并被认为是影响癌症的进程、预后和生存率的独立标志物。CHD1L活化的潜在机制可能是通过与Parp1、凋亡蛋白Nur77结合破坏细胞死亡程序,或者通过介导目的基因ARHGEF9、TCTP、SPOCK1、NTKL发挥相应功能。文中主要介绍了CHD1L在癌症中的作用和相关分子机制的研究进展。展开更多
Chromodomain-helicase-DNA-binding protein 1(CHD1)deletion is among the most common mutations in prostate cancer(PCa),but its role remains unclear.In this study,RNA sequencing was conducted in PCa cells after clustered...Chromodomain-helicase-DNA-binding protein 1(CHD1)deletion is among the most common mutations in prostate cancer(PCa),but its role remains unclear.In this study,RNA sequencing was conducted in PCa cells after clustered regularly interspaced palindromic repeat(CRISPR)/CRISPR-associated protein 9(Cas9)-based CHD1 knockout.Gene set enrichment analysis(GSEA)indicated upregulation of hypoxia-related pathways.A subsequent study confirmed that CHD1 deletion significantly upregulated hypoxia-inducible factor 1α(HIF1α)expression.Mechanistic investigation revealed that CHD1 deletion upregulated HIF1αby transcriptionally downregulating prolyl hydroxylase domain protein 2(PHD2),a prolyl hydroxylase catalyzing the hydroxylation of HIF1αand thus promoting its degradation by the E3 ligase von Hippel–Lindau tumor suppressor(VHL).Functional analysis showed that CHD1 deletion promoted angiogenesis and glycolysis,possibly through HIF1αtarget genes.Taken together,these findings indicate that CHD1 deletion enhances HIF1αexpression through PHD2 downregulation and therefore promotes angiogenesis and metabolic reprogramming in PCa.展开更多
Nucleosomes are the fundamental unit of chromatin.Chromatin remodeler plays a crucial role in the regulation of gene expression in eukaryotes.It is involved in important physiological processes,such as development,imm...Nucleosomes are the fundamental unit of chromatin.Chromatin remodeler plays a crucial role in the regulation of gene expression in eukaryotes.It is involved in important physiological processes,such as development,immune response,and metabolic regulation.During gene expression regulation,chromatin remodelers slide nucleosomes along genomic DNA and play a major role in chromatin organization.Chd1 senses the extranucleosomal linker DNA and controls nucleosome spacing in cells.However,the mechanism of linker DNA sensing by Chd1 is not completely understood.Here,we report the cryo-electron microscope(cryoEM)structures of Chd1 engaging nucleosomes in different states.Chd1 induces two exit-DNA conformations,either fully wrapped or partially unwrapped states.Notably,in the unwrapped conformation,the exit DNA interacts with a positively charged loop of the motor,named the exit-DNA binding loop,and traps Chd1 in the closed state in the ATPase cycle,suggesting attenuation of its remodeling activity.Explored single-molecule fluorescence resonance energy transfer(smFRET)and biochemical data supported the regulation of Chd1 remodeling activity by the exit-DNA conformations,which is important for the linker DNA sensitivity.Mutants of the Chd1 exit-DNA binding loop compromised nucleosome organization in yeast cells.Together,our findings provide valuable insights into Chd1 regulation by exit DNA unwrapping.These results provide a new perspective for the study of cell development and metabolism.展开更多
文摘目的:分析chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)基因在结直肠癌组织中的表达情况及CHD1L基因表达对结直肠癌细胞侵袭和转移能力的影响.方法:收集结直肠癌患者癌和癌旁组织,通过荧光定量PCR、免疫组织化学及Western blot检测CHD1L的表达情况,运用Transwell实验、划痕实验研究CHD1L对结直肠癌细胞侵袭和转移能力的影响.结果:CHD1L mRNA和蛋白在结直肠癌组织中表达水平显著高于与之对应的癌旁组织,差异有统计学意义;此外,结肠癌细胞株中CHD1L的表达普遍呈现高表达.结肠癌细胞LOVO和DLD-1中转染shRNA-CHD1L后,可以明显降低CHD1L的mRNA和蛋白的表达,同时明显抑制结肠癌细胞的侵袭和迁移能力.体内实验进一步发现降低SW620细胞中CHD1L的表达可以明显抑制肿瘤的肝、肺转移.结论:CHD1L在结直肠癌组织和细胞中呈现高表达,CHD1L在结直肠癌的侵袭和转移过程中发挥重要作用.CHD1L有可能成为治疗结直肠癌的新的靶点.
文摘目的:检测chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)在结直肠组织中的表达情况及探讨CHD1L基因对结直肠癌(colorectal cancer,CRC)细胞增殖和凋亡的影响.方法:通过荧光定量PCR、免疫组织化学检测80例CRC患者癌组织和对应癌旁组织中CHD1L基因的表达情况;采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)、克隆形成实验及动物实验等观察CHD1L基因在CRC细胞增殖中的作用;利用流式细胞技术检测细胞凋亡.结果:76.25%(61/80)的CRC癌组织CHD1L基因的表达明显高于对应的癌旁组织;转染s h R N A-C H D1L至C R C细胞可以明显降低C H D1L基因的m R N A和蛋白的表达,同时M T T实验和克隆形成实验发现干扰C H D1L组细胞的增殖明显低于对照组.另外,流式细胞仪检测发现干扰CHD1L组较对照组细胞凋亡指数明显增加.裸鼠成瘤实验发现稳定低表达CHD1L组肿瘤体积和重量明显小于对照组.结论:CHD1L在结直肠组织呈现高表达,降低结直肠细胞中CHD1L的表达可明显抑制细胞增殖及肿瘤生长.CHD1L可能成为治疗CRC的新靶点.
文摘Chromodomain helicase/ATPase DNA binding protein 1-like gene(CHD1L)是一种新发现的位于染色体1q21致癌基因,主要通过细胞增殖、G1/S期的转变、抑制凋亡、染色质不稳定等机制致癌,并被认为是影响癌症的进程、预后和生存率的独立标志物。CHD1L活化的潜在机制可能是通过与Parp1、凋亡蛋白Nur77结合破坏细胞死亡程序,或者通过介导目的基因ARHGEF9、TCTP、SPOCK1、NTKL发挥相应功能。文中主要介绍了CHD1L在癌症中的作用和相关分子机制的研究进展。
基金This study was supported by grants from the National Natural Science Foundation of China(No.82173292 and No.82002693).
文摘Chromodomain-helicase-DNA-binding protein 1(CHD1)deletion is among the most common mutations in prostate cancer(PCa),but its role remains unclear.In this study,RNA sequencing was conducted in PCa cells after clustered regularly interspaced palindromic repeat(CRISPR)/CRISPR-associated protein 9(Cas9)-based CHD1 knockout.Gene set enrichment analysis(GSEA)indicated upregulation of hypoxia-related pathways.A subsequent study confirmed that CHD1 deletion significantly upregulated hypoxia-inducible factor 1α(HIF1α)expression.Mechanistic investigation revealed that CHD1 deletion upregulated HIF1αby transcriptionally downregulating prolyl hydroxylase domain protein 2(PHD2),a prolyl hydroxylase catalyzing the hydroxylation of HIF1αand thus promoting its degradation by the E3 ligase von Hippel–Lindau tumor suppressor(VHL).Functional analysis showed that CHD1 deletion promoted angiogenesis and glycolysis,possibly through HIF1αtarget genes.Taken together,these findings indicate that CHD1 deletion enhances HIF1αexpression through PHD2 downregulation and therefore promotes angiogenesis and metabolic reprogramming in PCa.
基金supported by the National Natural Science Foundation of China(20211310041)the Beijing Municipal Natural Science Foundation(7244333)+1 种基金the Beijing Hospitals Authority’s Ascent Plan(DFL20241201)the Natural Science Foundation of Henan Province(232300421039).
文摘Nucleosomes are the fundamental unit of chromatin.Chromatin remodeler plays a crucial role in the regulation of gene expression in eukaryotes.It is involved in important physiological processes,such as development,immune response,and metabolic regulation.During gene expression regulation,chromatin remodelers slide nucleosomes along genomic DNA and play a major role in chromatin organization.Chd1 senses the extranucleosomal linker DNA and controls nucleosome spacing in cells.However,the mechanism of linker DNA sensing by Chd1 is not completely understood.Here,we report the cryo-electron microscope(cryoEM)structures of Chd1 engaging nucleosomes in different states.Chd1 induces two exit-DNA conformations,either fully wrapped or partially unwrapped states.Notably,in the unwrapped conformation,the exit DNA interacts with a positively charged loop of the motor,named the exit-DNA binding loop,and traps Chd1 in the closed state in the ATPase cycle,suggesting attenuation of its remodeling activity.Explored single-molecule fluorescence resonance energy transfer(smFRET)and biochemical data supported the regulation of Chd1 remodeling activity by the exit-DNA conformations,which is important for the linker DNA sensitivity.Mutants of the Chd1 exit-DNA binding loop compromised nucleosome organization in yeast cells.Together,our findings provide valuable insights into Chd1 regulation by exit DNA unwrapping.These results provide a new perspective for the study of cell development and metabolism.
