Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.How...Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.However,relatively few studies have explored the impact of post-translational modifications(PTM)on OC progression,which is essential for uncovering new therapeutic targets.This study aimed to systematically identify the key PTM types involved in OCprogression,and to explore and evaluate their translational potential as therapeutic targets.Methods:First,we utilized multiple general PTM antibodies to compare gross PTM levels between normal ovarian and OC tissues from clinical females.After identifying lactylation as the PTM with the most significant differences,we selected representative samples for label-free mass spectrometry to identify specific lactylation sites.Next,we transfected A2780(OC)cells with either wild-type(WT)or mutant(K192A[Q])poly(ADP-ribose)polymerase 1(PARP1)conjugated to enhanced green fluorescent protein(EGFP)with a StrepⅡpeptide tag and assessed various cellular indexes related to cell proliferation(clonogenicity assay),migration(scratch wound healing assay),and reactive oxygen species levels.Results:Pan-lactylation was significantly upregulated in clinical OC samples,with PARP1 lactylation at K192 being one of the most common modifications.The growth and migration of A2780 cells were markedly suppressed by overexpressing PARP1-WT but not mutant PARP1.Overexpressing PARP1 significantly downregulated the phosphorylation of extracellular signal-regulated kinases 1/2(ERK1/2).Conclusion:This study uncovered a novel PTM of PARP1 in OC,lactylation,and demonstrated that lactylation at K192 is crucial in regulating OC cell growth and migration via the ERK1/2 pathway.Further investigations are required to elucidate the broader functional implications of PARP1 lactylation and its therapeutic potential.展开更多
PARP1和CDK2在癌症发生发展中起关键作用,二者过度活化与多种肿瘤相关,同时抑制二者活性可能为乳腺癌治疗提供有效策略。本文以PARP1抑制剂Olaparib和CDK抑制剂Ribociclib为基础,通过分子对接分析选择分子尾部游离基团作为改造位点,设...PARP1和CDK2在癌症发生发展中起关键作用,二者过度活化与多种肿瘤相关,同时抑制二者活性可能为乳腺癌治疗提供有效策略。本文以PARP1抑制剂Olaparib和CDK抑制剂Ribociclib为基础,通过分子对接分析选择分子尾部游离基团作为改造位点,设计合成了5个具有全新结构的PARP-CDK双靶点抑制剂。利用核磁共振氢谱对其结构进行表征,通过测试酶抑制活性和体外抗细胞增殖活性进行生物活性评价。结果显示,双杂环链接化合物活性优于单杂环化合物,其中链接链为3,9-二氮杂螺[5.5]十一烷的化合物12c对PARP1与CDK2显示出良好的抗肿瘤效果。本研究为开发更多PARP1双靶点抑制剂奠定了重要基础。The proteins PARP1 and CDK2 are pivotal in the progression of cancer, with their excessive activation being implicated in a wide range of tumors. Targeting and simultaneously inhibiting the activities of both PARP1 and CDK2 may hold promise as a highly effective therapeutic strategy for the treatment of breast cancer. In this paper, five PARP-CDK dual-target inhibitors with a novel structure were designed and synthesized based on the PARP1 inhibitor Olaparib and the CDK inhibitor Ribociclib. The molecular docking analysis was used to select the free groups at the molecular tail as the modification sites. The structures of the inhibitors were characterized by nuclear magnetic resonance spectroscopy, and their biological activities were evaluated by testing the enzyme inhibitory activity and in vitro anti-cell proliferation activity. The results showed that the activity of the compounds with double heterocyclic linkers was better than that of the single heterocyclic compounds, and the compound 12c with a 3,9-diazaspiro [5.5] undecane linker exhibited good antitumor effects against both PARP1 and CDK2. This study lays an important foundation for the development of more PARP1 dual-target inhibitors.展开更多
In this study,we reported the discovery and structure-activity relationship analysis of chrysin derivatives as a new class of inhibitors targeting poly(ADP-ribose)polymerase 1(PARP1).Among these derivatives,compound 5...In this study,we reported the discovery and structure-activity relationship analysis of chrysin derivatives as a new class of inhibitors targeting poly(ADP-ribose)polymerase 1(PARP1).Among these derivatives,compound 5d emerged as the most effective chrysin-based inhibitor of PARP1,with an IC50 value of 108 nmol·L^(-1).This compound significantly inhibited the proliferation and migration of breast cancer cell lines HCC-1937 and MDA-MB-436 by inducing DNA damage.Furthermore,5d induced apoptosis and caused an extended G1/S-phase in these cell lines.Molecular docking studies revealed that 5d possesses a strong binding affinity toward PARP1.In vivo,in a xenograft model,5d effectively reduced tumor growth by downregulating PARP1 expression.Overall,compound 5d shows promise as a potential therapeutic agent for the treatment of BRCA wild-type breast cancer.展开更多
Osteoarthritis(OA)is an age-related cartilage-degenerating joint disease.Mitochondrial dysfunction has been reported to promote the development of OA.Poly(ADP-ribose)polymerase family member 12(PARP12)is a key regulat...Osteoarthritis(OA)is an age-related cartilage-degenerating joint disease.Mitochondrial dysfunction has been reported to promote the development of OA.Poly(ADP-ribose)polymerase family member 12(PARP12)is a key regulator of mitochondrial function,protein translation,and inflammation.However,the role of PARP12 in OA-based cartilage degradation and the underlying mechanisms are relatively unknown.Here,we first demonstrated that PARP12 inhibits mitophagy and promotes OA progression in human OA cartilage and a monosodium iodoacetate-induced rat OA model.Using mass spectrometry and co-immunoprecipitation assay,PARP12 was shown to interact with ISG15,upregulate mitofusin 1 and 2(MFN1/2)ISGylation,which downregulated MFN1/2 ubiquitination and SUMOylation,thereby inhibiting PINK1/Parkin-dependent chondrocyte mitophagy and promoting cartilage degradation.Moreover,inflammatory cytokine-induced interferon regulatory factor 1(IRF1)activation was required for the upregulation of PARP12 expression,and it directly bound to the PARP12 promoter to activate transcription.XAV-939 inhibited PARP12 expression and suppressed OA pathogenesis in vitro and in vivo.Clinically,PARP12 can be used to predict the severity of OA;thus,it represents a new target for the study of mitophagy and OA progression.In brief,the IRF1-mediated upregulation of PARP12 promoted cartilage degradation by inhibiting PINK1/Parkin-dependent mitophagy via ISG15-based attenuation of MFN1/2 ubiquitylation and SUMOylation.Our data provide new insights into the molecular mechanisms underlying PARP12-based regulation of mitophagy and can facilitate the development of therapeutic strategies for the treatment of OA.展开更多
Objective:Alternative splicing affects gene expression during placental development.The present study aimed to identify poly(ADP-ribose)polymerase 1(PARP1)-regulated alternative splicing events in HTR-8/Svneo cells.Me...Objective:Alternative splicing affects gene expression during placental development.The present study aimed to identify poly(ADP-ribose)polymerase 1(PARP1)-regulated alternative splicing events in HTR-8/Svneo cells.Methods:Decidual tissues were collected from women with induced abortion and spontaneous abortion.PARP1 transcription was quantified by RT-qPCR.Small interfering RNA(siRNA)was used to knock down the PARP1 expression in HTR-8/Svneo cells.The transfection efficiency was verified by RT-qPCR and Western blotting.Total RNA was extracted,and the RNA-sequencing approach was used to identify alternative splicing events and transcriptomes.The PARP1 knockdown-induced differentially expressed genes with changes in alternative splicing events were quantified by RT-qPCR.Functional analysis,which included the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways,was performed.Results:The PARP1 mRNA expression increased in decidual tissues in the spontaneous abortion group,when compared to the induced abortion group.However,the PARP1 knockdown significantly downregulated 1491 genes and upregulated 881 genes in HTR-8/Svneo cells.Furthermore,227 genes that underwent alternative splicing were identified,and these were differentially expressed in siPARP1 cells,when compared to siNC cells.Conclusion:The functional analysis revealed that these alternative splicing genes affected the functional phenotypes of extravillous cytotrophoblasts.Furthermore,the PARP1 knockdown led to alterations in gene expression and specific alternative splicing patterns in extravillous trophoblasts.展开更多
文摘Objective:Ovarian cancer(OC)ranks among the leading causes of mortality among the female cancers worldwide.Numerous studies have explored the development and progression of OC at multiple genetic regulatory levels.However,relatively few studies have explored the impact of post-translational modifications(PTM)on OC progression,which is essential for uncovering new therapeutic targets.This study aimed to systematically identify the key PTM types involved in OCprogression,and to explore and evaluate their translational potential as therapeutic targets.Methods:First,we utilized multiple general PTM antibodies to compare gross PTM levels between normal ovarian and OC tissues from clinical females.After identifying lactylation as the PTM with the most significant differences,we selected representative samples for label-free mass spectrometry to identify specific lactylation sites.Next,we transfected A2780(OC)cells with either wild-type(WT)or mutant(K192A[Q])poly(ADP-ribose)polymerase 1(PARP1)conjugated to enhanced green fluorescent protein(EGFP)with a StrepⅡpeptide tag and assessed various cellular indexes related to cell proliferation(clonogenicity assay),migration(scratch wound healing assay),and reactive oxygen species levels.Results:Pan-lactylation was significantly upregulated in clinical OC samples,with PARP1 lactylation at K192 being one of the most common modifications.The growth and migration of A2780 cells were markedly suppressed by overexpressing PARP1-WT but not mutant PARP1.Overexpressing PARP1 significantly downregulated the phosphorylation of extracellular signal-regulated kinases 1/2(ERK1/2).Conclusion:This study uncovered a novel PTM of PARP1 in OC,lactylation,and demonstrated that lactylation at K192 is crucial in regulating OC cell growth and migration via the ERK1/2 pathway.Further investigations are required to elucidate the broader functional implications of PARP1 lactylation and its therapeutic potential.
文摘PARP1和CDK2在癌症发生发展中起关键作用,二者过度活化与多种肿瘤相关,同时抑制二者活性可能为乳腺癌治疗提供有效策略。本文以PARP1抑制剂Olaparib和CDK抑制剂Ribociclib为基础,通过分子对接分析选择分子尾部游离基团作为改造位点,设计合成了5个具有全新结构的PARP-CDK双靶点抑制剂。利用核磁共振氢谱对其结构进行表征,通过测试酶抑制活性和体外抗细胞增殖活性进行生物活性评价。结果显示,双杂环链接化合物活性优于单杂环化合物,其中链接链为3,9-二氮杂螺[5.5]十一烷的化合物12c对PARP1与CDK2显示出良好的抗肿瘤效果。本研究为开发更多PARP1双靶点抑制剂奠定了重要基础。The proteins PARP1 and CDK2 are pivotal in the progression of cancer, with their excessive activation being implicated in a wide range of tumors. Targeting and simultaneously inhibiting the activities of both PARP1 and CDK2 may hold promise as a highly effective therapeutic strategy for the treatment of breast cancer. In this paper, five PARP-CDK dual-target inhibitors with a novel structure were designed and synthesized based on the PARP1 inhibitor Olaparib and the CDK inhibitor Ribociclib. The molecular docking analysis was used to select the free groups at the molecular tail as the modification sites. The structures of the inhibitors were characterized by nuclear magnetic resonance spectroscopy, and their biological activities were evaluated by testing the enzyme inhibitory activity and in vitro anti-cell proliferation activity. The results showed that the activity of the compounds with double heterocyclic linkers was better than that of the single heterocyclic compounds, and the compound 12c with a 3,9-diazaspiro [5.5] undecane linker exhibited good antitumor effects against both PARP1 and CDK2. This study lays an important foundation for the development of more PARP1 dual-target inhibitors.
基金This work was supported by the Clinical Research Center for Breast&Thyroid Disease Prevention in Hunan Province(No.2018SK4001)the Scientific Research Project of Hunan Provincial Health Commission(No.20201969)。
文摘In this study,we reported the discovery and structure-activity relationship analysis of chrysin derivatives as a new class of inhibitors targeting poly(ADP-ribose)polymerase 1(PARP1).Among these derivatives,compound 5d emerged as the most effective chrysin-based inhibitor of PARP1,with an IC50 value of 108 nmol·L^(-1).This compound significantly inhibited the proliferation and migration of breast cancer cell lines HCC-1937 and MDA-MB-436 by inducing DNA damage.Furthermore,5d induced apoptosis and caused an extended G1/S-phase in these cell lines.Molecular docking studies revealed that 5d possesses a strong binding affinity toward PARP1.In vivo,in a xenograft model,5d effectively reduced tumor growth by downregulating PARP1 expression.Overall,compound 5d shows promise as a potential therapeutic agent for the treatment of BRCA wild-type breast cancer.
基金supported by the National Natural Science Foundation of China (82202739)Guangdong Basic and Applied Basic Research Foundation Natural Science Fund Project-Youth Promotion Project (2024A1515030260)+1 种基金China Statefunded Postdoctoral Researchers Program (GZC20233265)National Natural Science Foundation of China (81972051)。
文摘Osteoarthritis(OA)is an age-related cartilage-degenerating joint disease.Mitochondrial dysfunction has been reported to promote the development of OA.Poly(ADP-ribose)polymerase family member 12(PARP12)is a key regulator of mitochondrial function,protein translation,and inflammation.However,the role of PARP12 in OA-based cartilage degradation and the underlying mechanisms are relatively unknown.Here,we first demonstrated that PARP12 inhibits mitophagy and promotes OA progression in human OA cartilage and a monosodium iodoacetate-induced rat OA model.Using mass spectrometry and co-immunoprecipitation assay,PARP12 was shown to interact with ISG15,upregulate mitofusin 1 and 2(MFN1/2)ISGylation,which downregulated MFN1/2 ubiquitination and SUMOylation,thereby inhibiting PINK1/Parkin-dependent chondrocyte mitophagy and promoting cartilage degradation.Moreover,inflammatory cytokine-induced interferon regulatory factor 1(IRF1)activation was required for the upregulation of PARP12 expression,and it directly bound to the PARP12 promoter to activate transcription.XAV-939 inhibited PARP12 expression and suppressed OA pathogenesis in vitro and in vivo.Clinically,PARP12 can be used to predict the severity of OA;thus,it represents a new target for the study of mitophagy and OA progression.In brief,the IRF1-mediated upregulation of PARP12 promoted cartilage degradation by inhibiting PINK1/Parkin-dependent mitophagy via ISG15-based attenuation of MFN1/2 ubiquitylation and SUMOylation.Our data provide new insights into the molecular mechanisms underlying PARP12-based regulation of mitophagy and can facilitate the development of therapeutic strategies for the treatment of OA.
基金supported by the State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia Fund(No.SKL-HIDCA-2020-JZ11).
文摘Objective:Alternative splicing affects gene expression during placental development.The present study aimed to identify poly(ADP-ribose)polymerase 1(PARP1)-regulated alternative splicing events in HTR-8/Svneo cells.Methods:Decidual tissues were collected from women with induced abortion and spontaneous abortion.PARP1 transcription was quantified by RT-qPCR.Small interfering RNA(siRNA)was used to knock down the PARP1 expression in HTR-8/Svneo cells.The transfection efficiency was verified by RT-qPCR and Western blotting.Total RNA was extracted,and the RNA-sequencing approach was used to identify alternative splicing events and transcriptomes.The PARP1 knockdown-induced differentially expressed genes with changes in alternative splicing events were quantified by RT-qPCR.Functional analysis,which included the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways,was performed.Results:The PARP1 mRNA expression increased in decidual tissues in the spontaneous abortion group,when compared to the induced abortion group.However,the PARP1 knockdown significantly downregulated 1491 genes and upregulated 881 genes in HTR-8/Svneo cells.Furthermore,227 genes that underwent alternative splicing were identified,and these were differentially expressed in siPARP1 cells,when compared to siNC cells.Conclusion:The functional analysis revealed that these alternative splicing genes affected the functional phenotypes of extravillous cytotrophoblasts.Furthermore,the PARP1 knockdown led to alterations in gene expression and specific alternative splicing patterns in extravillous trophoblasts.
