目的:研究CLN3基因在卵巢浆液性癌及子宫内膜癌中的表达特点及与其发生发展的关系。方法:采用RT-PCR和W estern B lot方法检测卵巢浆液性肿瘤(良性浆液性肿瘤、交界性肿瘤、浆液性癌)组织、子宫内膜癌及癌旁组织、正常卵巢组织和正常子...目的:研究CLN3基因在卵巢浆液性癌及子宫内膜癌中的表达特点及与其发生发展的关系。方法:采用RT-PCR和W estern B lot方法检测卵巢浆液性肿瘤(良性浆液性肿瘤、交界性肿瘤、浆液性癌)组织、子宫内膜癌及癌旁组织、正常卵巢组织和正常子宫内膜组织中CLN3基因表达情况。结果:卵巢浆液性癌组织中CLN3 mRNA表达为3.04±0.58,交界性卵巢浆液性肿瘤组织中表达为1.63±0.43,均明显高于正常卵巢组织和良性卵巢浆液性肿瘤,差异显著(P<0.01);卵巢浆液性癌组织CLN3表达高于交界性卵巢浆液性肿瘤(P<0.01);卵巢浆液性癌患者不同临床分期(Ⅰ/Ⅱ期与Ⅲ/Ⅳ期)、病理分级及有无腹水间比较差异显著(P<0.05)。正常子宫内膜组织中CLN3呈低表达,在所有的子宫内膜癌及癌旁组织中发现CLN3 mRNA及蛋白表达上调(P<0.01)。结论:①CLN3基因与卵巢浆液性癌的发生发展、生物学行为和预后可能有关,可作为卵巢浆液性癌的重要的生物学标记物,是卵巢浆液癌防治的一个新靶点;②CLN3基因过表达可能与子宫内膜癌发生有关。展开更多
Background The fat deposition has a crucial role in animal meat flavor,and fat deposition-related traits are vital for breeding in the commercial duck industry.Avian fat-related traits are typical complex phenotypes,w...Background The fat deposition has a crucial role in animal meat flavor,and fat deposition-related traits are vital for breeding in the commercial duck industry.Avian fat-related traits are typical complex phenotypes,which need a large amount of data to analyze the genetic loci.Results In this study,we performed a new phenotypic analysis of fat traits and genotyped whole-genome varia-tions for 1,246 ducks,and combed with previous GWAS data to reach 1,880 ducks for following analysis.The carcass composition traits,subcutaneous fat weight(SFW),subcutaneous fat percentage(SFP),abdominal fat weight(AFW),abdominal fat percentage(AFP)and the body weight of day 42(BW42)for each duck were collected.We identified a set of new loci that affect the traits related to fat deposition in avian.Among these loci,ceroid-lipofuscinosis,neuronal 8(CLN8)is a novel candidate gene controlling fat deposition.We investigated its novel function and regulation in avian adipogenesis.Five significant SNPs(the most significant SNP,P-value=21.37E-12)and a single haplotype were detected in the upstream of CLN8 for subcutaneous fat percentage.Subsequently,luciferase assay demonstrated that 5 linked SNPs in the upstream of the CLN8 gene significantly decreased the transcriptional activity of CLN8.Further,ATAC-seq analysis showed that transcription factor binding sites were identified in a region close to the haplotype.A set of luciferase reporter gene vectors that contained different deletion fragments of the CLN8 promoter were con-structed,and the core promoter area of CLN8 was finally identified in the-1,884/-1,207 bp region of the 5′flanking sequences,which contains adipogenesis-related transcription factors binding sites.Moreover,the over-expression of CLN8 can remarkably facilitate adipocyte differentiation in ICPs.Consistent with these,the global transcriptome profiling and functional analysis of the over-expressed CLN8 in the cell line further revealed that the lipid biosynthetic process during the adipogenesis was significantly enriched.Conclusions Our results demonstrated that CLN8 is a positive regulator of avian adipocyte differentiation.These findings identify a novel function of CLN8 in adipocyte differentiation,which provides important clues for the further study of the mechanism of avian fat deposition.展开更多
目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的...目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的方法检测CLN3在卵巢浆液性囊腺癌中的表达,分析其表达与临床病理特征及化疗耐药的相关性。结果 CLN3在卵巢浆液性囊腺癌中的表达高于正常卵巢组织,且临床分期越晚其表达越高(P<0.05),而与肿瘤患者年龄无显著相关性。CLN3在卵巢癌化疗耐药组表达阳性率明显高于化疗敏感组(P<0.05)。结论卵巢癌中CLN3高表达与卵巢癌的发生、发展、浸润和转移有关,与卵巢癌化疗耐药密切相关。CLN3可成为卵巢癌化疗新的基因治疗靶点。展开更多
BACKGROUND Adult neuronal ceroid lipofuscinosis(ANCL)can be caused by compound heterozygous recessive mutations in CLN6.The main clinical features of the disease are neurodegeneration,progressive motor dysfunction,sei...BACKGROUND Adult neuronal ceroid lipofuscinosis(ANCL)can be caused by compound heterozygous recessive mutations in CLN6.The main clinical features of the disease are neurodegeneration,progressive motor dysfunction,seizures,cognitive decline,ataxia,vision loss and premature death.CASE SUMMARY A 37-year-old female presented to our clinic with a 3-year history of limb weakness and gradually experiencing unstable walking.The patient was diagnosed with CLN6 type ANCL after the identification of mutations in the CLN6 gene.The patient was treated with antiepileptic drugs.The patient is under ongoing followup.Unfortunately,the patient’s condition has deteriorated,and she is currently unable to care for herself.CONCLUSION There is presently no effective treatment for ANCL.However,early diagnosis and symptomatic treatment are possible.展开更多
Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural pro...Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.展开更多
基金supported by the National Key R&D Program of China(2022YFF1000100)National Waterfowl-Industry Technology Research System(CARS-42–09)+1 种基金National Nature Science Foundation of China(31972525,31572388)Beijing Joint Research Program for Germplasm Innovation and New Variety Breeding(G20220628007).
文摘Background The fat deposition has a crucial role in animal meat flavor,and fat deposition-related traits are vital for breeding in the commercial duck industry.Avian fat-related traits are typical complex phenotypes,which need a large amount of data to analyze the genetic loci.Results In this study,we performed a new phenotypic analysis of fat traits and genotyped whole-genome varia-tions for 1,246 ducks,and combed with previous GWAS data to reach 1,880 ducks for following analysis.The carcass composition traits,subcutaneous fat weight(SFW),subcutaneous fat percentage(SFP),abdominal fat weight(AFW),abdominal fat percentage(AFP)and the body weight of day 42(BW42)for each duck were collected.We identified a set of new loci that affect the traits related to fat deposition in avian.Among these loci,ceroid-lipofuscinosis,neuronal 8(CLN8)is a novel candidate gene controlling fat deposition.We investigated its novel function and regulation in avian adipogenesis.Five significant SNPs(the most significant SNP,P-value=21.37E-12)and a single haplotype were detected in the upstream of CLN8 for subcutaneous fat percentage.Subsequently,luciferase assay demonstrated that 5 linked SNPs in the upstream of the CLN8 gene significantly decreased the transcriptional activity of CLN8.Further,ATAC-seq analysis showed that transcription factor binding sites were identified in a region close to the haplotype.A set of luciferase reporter gene vectors that contained different deletion fragments of the CLN8 promoter were con-structed,and the core promoter area of CLN8 was finally identified in the-1,884/-1,207 bp region of the 5′flanking sequences,which contains adipogenesis-related transcription factors binding sites.Moreover,the over-expression of CLN8 can remarkably facilitate adipocyte differentiation in ICPs.Consistent with these,the global transcriptome profiling and functional analysis of the over-expressed CLN8 in the cell line further revealed that the lipid biosynthetic process during the adipogenesis was significantly enriched.Conclusions Our results demonstrated that CLN8 is a positive regulator of avian adipocyte differentiation.These findings identify a novel function of CLN8 in adipocyte differentiation,which provides important clues for the further study of the mechanism of avian fat deposition.
文摘目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的方法检测CLN3在卵巢浆液性囊腺癌中的表达,分析其表达与临床病理特征及化疗耐药的相关性。结果 CLN3在卵巢浆液性囊腺癌中的表达高于正常卵巢组织,且临床分期越晚其表达越高(P<0.05),而与肿瘤患者年龄无显著相关性。CLN3在卵巢癌化疗耐药组表达阳性率明显高于化疗敏感组(P<0.05)。结论卵巢癌中CLN3高表达与卵巢癌的发生、发展、浸润和转移有关,与卵巢癌化疗耐药密切相关。CLN3可成为卵巢癌化疗新的基因治疗靶点。
文摘BACKGROUND Adult neuronal ceroid lipofuscinosis(ANCL)can be caused by compound heterozygous recessive mutations in CLN6.The main clinical features of the disease are neurodegeneration,progressive motor dysfunction,seizures,cognitive decline,ataxia,vision loss and premature death.CASE SUMMARY A 37-year-old female presented to our clinic with a 3-year history of limb weakness and gradually experiencing unstable walking.The patient was diagnosed with CLN6 type ANCL after the identification of mutations in the CLN6 gene.The patient was treated with antiepileptic drugs.The patient is under ongoing followup.Unfortunately,the patient’s condition has deteriorated,and she is currently unable to care for herself.CONCLUSION There is presently no effective treatment for ANCL.However,early diagnosis and symptomatic treatment are possible.
文摘Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.
