Genomic destabilization and defective DNA repair are the most prominent features of tumour cells and are exploited by various chemotherapy drugs for cancer therapy.Long non-coding RNA(lncR-NAs)have emerged as powerful...Genomic destabilization and defective DNA repair are the most prominent features of tumour cells and are exploited by various chemotherapy drugs for cancer therapy.Long non-coding RNA(lncR-NAs)have emerged as powerful regulators of gene expression and are thus involved in diverse biological processes.Recent studies have demonstrated that several lncRNAs play critical roles in DNA repair.Nonetheless,the relationship between DNA damage-responsive lncRNAs and chemoresistance remains poorly defined.In this study,we established four different DNA damage models triggered by cisplatin(DDP),H2O2,neocarzinostatin(NCS)or ultraviolet(UV)irradiation and identified a specific upregu-lated lncRNA(lnc-DUSP6)involved in the cisplatin-induced DNA damage response.Furthermore,loss-or gain-of-function experiments confirmed that lnc-DUSP6 enhanced DNA repair and cell survival under cisplatin treatment,thus promoting cisplatin resistance.Mechanistically,an RNA immunoprecipitation(RIP)assay revealed that lnc-DUSP6 directly interacts with DUSP6(Dual Specificity Phosphatase 6),which is closely associated with cisplatin sensitivity.Additionally,overexpression of DUSP6 significantly rescued the effects of lnc-DUSP6 silencing on DNA repair and cell survival under cisplatin treatment.O-verall,our results show the effect and underlying mechanism of lnc-DUSP6 in cisplatin resistance:lnc-DUSP6 promotes cisplatin-induced DNA damage repair and cisplatin resistance by stabilizing DUSP6,which is highly clinically important for enhancing the efficacy of cisplatin for cancers.展开更多
Objective Cytokine responses to activation of innate immunity differ between individuals,yet the genomic and tissue-specific transcriptomic determinants of inflammatory responsiveness are not well understood. We hypot...Objective Cytokine responses to activation of innate immunity differ between individuals,yet the genomic and tissue-specific transcriptomic determinants of inflammatory responsiveness are not well understood. We hypothesized that tissue-specific mRNA and long intergenic non-coding RNA (lincRNA) induction differs between individuals with divergent evoked inflammatory responses.展开更多
Vascular remodeling is the essential pathogenic process of various cardiovascular disorders,including hypertension,atherosclerosis,stroke,and restenosis after vein graft.The main characterization of vascular remodelin...Vascular remodeling is the essential pathogenic process of various cardiovascular disorders,including hypertension,atherosclerosis,stroke,and restenosis after vein graft.The main characterization of vascular remodeling is abnormal variations of vascular cell phenotype,morphological structure and functions such as migration,hypertrophy,proliferation and apoptosis.Numerous researches revealed that mechanical stress,including shear stress and cyclic stretch,participates in physiological vascular homeostasis,or pathophysiological vascular remodeling.The understanding of mechanobiological mechanism in vascular remodeling will play a unique role in understanding human physiology and disease,and will generate important theoretical and clinical significance [2].Non-coding RNAs are newly recognized RNAs which cannot be translated into proteins but are involved in epigenetic modification of gene regulation.The studies revealed that non-coding RNAs,such as microRNAs(miRNAs)and long noncoding RNAs(long ncRNAs,IncRNA),as well as small interfering RNAs(siRNAs),piwi-interacting RNAs(piRNAs),small nucleolar RNAs(snoRNAs),play essential roles in the regulation of various processes,such as metabolism,development,cell proliferation,cell apoptosis,cell differentiation,oncogenesis and vascular homeostasis[5].However,the roles of non-coding RNAs in the cardiovascular system under mechanical stresses are still not clarified.Our recent researches detected the mechanical regulation of IncRNAs and miRNAs in vascular remodeling.LncRNAs are non-protein-coding transcripts that are longer than 200 nucleotides(nt),which is an arbitrary cut-off value that distinguishes these transcripts from other small RNAs.Unlike the well-established mechanism of microRNA action,the functional mode of IncRNAs is not fully understood.Increasing evidence shows that IncRNAs modulate gene expression via a multilevel-regulated pathway.Given their large number and complicated functional modes,lncRNAs are emerging as important regulators of a variety of cellular responses,developmental processes and diseases.Using a gene microarray,we screened the differences in the IncRNAs and mRNAs between spontaneously hypertensive rats(SHR)and Wistar Kyoto rats(WKY).The results showed that 68 IncRNAs and 255 mRNAs were up-regulated in the aorta of SHR,while 167 IncRNAs and 272 mRNAs were down-regulated.Expressions of the screened IncRNAs,including XR007793,were validated by real-time PCR.A co-expression network was composed,and gene function was analysed using Ingenuity Pathway Analysis.In vitro,vascular smooth muscle cells(VSMCs)were subjected to cyclic stretch at a magnitude of 5%(physiological normotensive cyclic stretch)or 15%(pathological hypertensive cyclic stretch)by Flexercell-5000TM.15%-cyclic-stretch increased XR007793 expression.XR007793 knockdown attenuated VSMC proliferation and migration and inhibited co-expressed genes such as signal transducers and activators of transcription 2(stat2),LIM domain only 2(lmo2)and interferon regulatory factor 7(irf7)[4].Illuminating the role of IncRNAs in vascular remodeling induced by hyper mechanical stretch may provide deeper insight into the mechanobiological mechanism underlying hypertension,and contribute to identifying potential targets for hypertension therapy.miRNAs are endogenous,non-coding,single-stranded RNAs of 18-22 nucleotides that constitute a novel class of gene regulators.miRNAs bind to their target genes within their 3’-untranslated regions(3’-UTRs),leading to direct degradation of mRNA or translational repression by a complete,i.e.in plants,or incomplete,i.e.in animals,complement respectively.Our resent works revealed several important mechano-responsive miRNA and their potential effects in vascular remodeling.Forexample,miRNA-33 is regulated by cyclic stretch in the grafted vessels,which targets to BMP3 and subsequent modulates smad signaling pathway.The miRNA-33-BMP3-smad pathway protects against venous VSMC proliferation in response to arterial cyclic stretch.Therefore,miRNA-33 may be a potential therapeutic target in autologous vein grafted surgery,and locally overexpression of miR-33 may attenuates neointimal hyperplasia of grafted human saphenous vein [3].The unpublished data revealed that 15%cyclic stretch also significantly elevated the expression of miRNA-124-3p which bound to the 3’UTR of Lmna mRNA,and then negatively regulated protein expression of lamin A/C which is the important skeletal proteins in nucleus.In addition to primary intracellular locations of miRNAs,our recent study showed that miRNAs can be secreted and protected extracellularly via inclusion into membrane-derived vesicles including microparticles.Microparticles are extracellular vesicles ranging from 0.1 to 1μm in size and have been shown to deliver various bioactive molecules,i.e.,chemokines,enzymes and miRNAs,to recipient cells.Increasing evidence shows that microparticles play a pivotal role in many pathological processes,such as cancer,inflammatory diseases and cardiovascular disease.Our present study showed that platelet-derived microparticles(PMPs),which are released by active platelets,are important vehicles for communication and play crucial roles in inducing abnormal EC proliferation in hypertension.In briefly,EC proliferation was increased in renal hypertensive rats established by abdominal aortic coarctation compared to control rats and that elevated thrombin in plasma promoted platelet activation,which may induce the release of PMPs.miRNA array and qPCR revealed a higher level of miRNA-142-3p in platelets and PMPs.In vitro,PMPs delivered miRNA-142-3p into ECs and enhanced EC proliferation via Bcl-2-associated transcription factor 1(BCLAF1)and its downstream genes.These results indicated that PMPs deliver miRNA-142-3p from activated platelets into ECs and that miRNA-142-3p may play important roles in EC dysfunction under hypertensive conditions and might be a novel therapeutic target for maintaining EC homeostasis in hypertension[1].These results provide possible mechanisms by which non-coding RNAs regulate cellular functions under different mechanical stresses,and suggest a novel potential therapeutic approach for vascular remodeling.The further studies on noncoding RNAs may provide new insight into understanding the mechanism of vascular remodeling in different various cardiovascular disorders,and may provide novel targets for the maintenance of vascular homeostasis.展开更多
以高耐热性玉米品种郑单958、低耐热性玉米品种先玉335为试验材料,以正常生长条件为对照(CK),利用半自动伸缩高温棚进行花期高温胁迫(HT)处理,通过circRNA高通量测序筛选高温胁迫下不同玉米品种花粉中差异表达的环状RNA(circRNA),对其...以高耐热性玉米品种郑单958、低耐热性玉米品种先玉335为试验材料,以正常生长条件为对照(CK),利用半自动伸缩高温棚进行花期高温胁迫(HT)处理,通过circRNA高通量测序筛选高温胁迫下不同玉米品种花粉中差异表达的环状RNA(circRNA),对其来源基因进行GO和KEGG富集分析,并筛选具有miRNA结合位点的差异表达circRNA,预测其下游目的基因,分析玉米花粉中响应高温胁迫的潜在circRNA-miRNA-mRNA共表达调控网络,从多层面解析玉米花粉中调控高温胁迫的分子作用机制,为提高玉米品种的耐热性提供理论依据。结果表明,在郑单958、先玉335不同样本中共鉴定出1 843个不同的circRNA,它们在玉米染色体中的分布不同。每个circRNA所包含的外显子数目也不相同,其中,大多数(624个)circRNA只含有1个外显子。在郑单958花粉中共鉴定出1 563个circRNA,其中,CK958-1、CK958-2、CK958-3中分别鉴定出305、213、356个circRNA,HT958-1、HT958-2、HT958-3中分别鉴定出222、242、225个circRNA。在先玉335花粉中共鉴定出1 423个circRNA,其中,CK335-1、CK335-2、CK335-3中分别鉴定出272、188、229个circRNA,HT335-1、HT335-2、HT335-3中分别鉴定出259、237、238个circRNA。不同样本中占比最高的均为外显子circRNA。circRNA与其来源基因不是一一对应的关系,有748个circRNA来源基因通过反向剪接机制只形成1个circRNA,156个circRNA来源基因通过反向剪接机制各自形成2个circRNA。在郑单958高温胁迫花粉与对照花粉对比组(HT958 vs CK958)中共筛选到9个差异表达circRNA,其中2个circRNA呈上调表达,其来源基因显著富集到焦磷酸酶活性、核苷酸磷酸代谢过程、糖基磷脂酰肌醇(GPI)锚定代谢过程等17个GO条目,显著富集到GPI锚定生物合成、代谢途径等KEGG通路。在先玉335高温胁迫花粉与对照花粉对比组(HT335 vs CK335)中共筛选到1个差异表达circRNA,其来源基因没有显著富集到任何GO条目、KEGG通路。在郑单958高温胁迫花粉与先玉335高温胁迫花粉对比组(HT958 vs HT335)中共筛选到17个差异表达circRNA,其中6个circRNA呈上调表达,其来源基因显著富集到内质网系统、高尔基相关囊泡膜、膜蛋白水解等16个GO条目中,没有显著富集到任何KEGG代谢通路。5个circRNA具有miRNA结合位点,可以作为海绵岛吸附miRNA间接调控下游靶标基因的表达,构建了包括5个circRNA、5个不同家族miRNA、2个mRNA在内的circRNA-miRNA-mRNA共表达调控网络。筛选到了54个circRNA包含内部核糖体进入位点(IRES),可以翻译表达多肽或者蛋白质直接作用于靶标基因。展开更多
基金Supported by the National Natural Science Foundation of China(No.82071571)the Natural Science Foundation of Guangdong Province(No.2021A1515010601)+3 种基金Guangdong Provincial Basic and Applied Basic Research Foundation-Dongguan Joint Fund(No.2024A1515140121)the“Climbing”Program of Guangdong Province(No.pdjh2021b0226)the Innovation and Entrepreneurship Program for College Students(No.GDMU2022038,202310571038,ZZDC002,S202510571041)Guangdong Medical University Undergraduate Innovation and Entrepreneurship Education Base Project(No.JDXM2024039,JDXM2025046)。
文摘Genomic destabilization and defective DNA repair are the most prominent features of tumour cells and are exploited by various chemotherapy drugs for cancer therapy.Long non-coding RNA(lncR-NAs)have emerged as powerful regulators of gene expression and are thus involved in diverse biological processes.Recent studies have demonstrated that several lncRNAs play critical roles in DNA repair.Nonetheless,the relationship between DNA damage-responsive lncRNAs and chemoresistance remains poorly defined.In this study,we established four different DNA damage models triggered by cisplatin(DDP),H2O2,neocarzinostatin(NCS)or ultraviolet(UV)irradiation and identified a specific upregu-lated lncRNA(lnc-DUSP6)involved in the cisplatin-induced DNA damage response.Furthermore,loss-or gain-of-function experiments confirmed that lnc-DUSP6 enhanced DNA repair and cell survival under cisplatin treatment,thus promoting cisplatin resistance.Mechanistically,an RNA immunoprecipitation(RIP)assay revealed that lnc-DUSP6 directly interacts with DUSP6(Dual Specificity Phosphatase 6),which is closely associated with cisplatin sensitivity.Additionally,overexpression of DUSP6 significantly rescued the effects of lnc-DUSP6 silencing on DNA repair and cell survival under cisplatin treatment.O-verall,our results show the effect and underlying mechanism of lnc-DUSP6 in cisplatin resistance:lnc-DUSP6 promotes cisplatin-induced DNA damage repair and cisplatin resistance by stabilizing DUSP6,which is highly clinically important for enhancing the efficacy of cisplatin for cancers.
文摘Objective Cytokine responses to activation of innate immunity differ between individuals,yet the genomic and tissue-specific transcriptomic determinants of inflammatory responsiveness are not well understood. We hypothesized that tissue-specific mRNA and long intergenic non-coding RNA (lincRNA) induction differs between individuals with divergent evoked inflammatory responses.
基金supported by grants from the National Natural Science Foundation of China ( 11625209,11572199,31670958)
文摘Vascular remodeling is the essential pathogenic process of various cardiovascular disorders,including hypertension,atherosclerosis,stroke,and restenosis after vein graft.The main characterization of vascular remodeling is abnormal variations of vascular cell phenotype,morphological structure and functions such as migration,hypertrophy,proliferation and apoptosis.Numerous researches revealed that mechanical stress,including shear stress and cyclic stretch,participates in physiological vascular homeostasis,or pathophysiological vascular remodeling.The understanding of mechanobiological mechanism in vascular remodeling will play a unique role in understanding human physiology and disease,and will generate important theoretical and clinical significance [2].Non-coding RNAs are newly recognized RNAs which cannot be translated into proteins but are involved in epigenetic modification of gene regulation.The studies revealed that non-coding RNAs,such as microRNAs(miRNAs)and long noncoding RNAs(long ncRNAs,IncRNA),as well as small interfering RNAs(siRNAs),piwi-interacting RNAs(piRNAs),small nucleolar RNAs(snoRNAs),play essential roles in the regulation of various processes,such as metabolism,development,cell proliferation,cell apoptosis,cell differentiation,oncogenesis and vascular homeostasis[5].However,the roles of non-coding RNAs in the cardiovascular system under mechanical stresses are still not clarified.Our recent researches detected the mechanical regulation of IncRNAs and miRNAs in vascular remodeling.LncRNAs are non-protein-coding transcripts that are longer than 200 nucleotides(nt),which is an arbitrary cut-off value that distinguishes these transcripts from other small RNAs.Unlike the well-established mechanism of microRNA action,the functional mode of IncRNAs is not fully understood.Increasing evidence shows that IncRNAs modulate gene expression via a multilevel-regulated pathway.Given their large number and complicated functional modes,lncRNAs are emerging as important regulators of a variety of cellular responses,developmental processes and diseases.Using a gene microarray,we screened the differences in the IncRNAs and mRNAs between spontaneously hypertensive rats(SHR)and Wistar Kyoto rats(WKY).The results showed that 68 IncRNAs and 255 mRNAs were up-regulated in the aorta of SHR,while 167 IncRNAs and 272 mRNAs were down-regulated.Expressions of the screened IncRNAs,including XR007793,were validated by real-time PCR.A co-expression network was composed,and gene function was analysed using Ingenuity Pathway Analysis.In vitro,vascular smooth muscle cells(VSMCs)were subjected to cyclic stretch at a magnitude of 5%(physiological normotensive cyclic stretch)or 15%(pathological hypertensive cyclic stretch)by Flexercell-5000TM.15%-cyclic-stretch increased XR007793 expression.XR007793 knockdown attenuated VSMC proliferation and migration and inhibited co-expressed genes such as signal transducers and activators of transcription 2(stat2),LIM domain only 2(lmo2)and interferon regulatory factor 7(irf7)[4].Illuminating the role of IncRNAs in vascular remodeling induced by hyper mechanical stretch may provide deeper insight into the mechanobiological mechanism underlying hypertension,and contribute to identifying potential targets for hypertension therapy.miRNAs are endogenous,non-coding,single-stranded RNAs of 18-22 nucleotides that constitute a novel class of gene regulators.miRNAs bind to their target genes within their 3’-untranslated regions(3’-UTRs),leading to direct degradation of mRNA or translational repression by a complete,i.e.in plants,or incomplete,i.e.in animals,complement respectively.Our resent works revealed several important mechano-responsive miRNA and their potential effects in vascular remodeling.Forexample,miRNA-33 is regulated by cyclic stretch in the grafted vessels,which targets to BMP3 and subsequent modulates smad signaling pathway.The miRNA-33-BMP3-smad pathway protects against venous VSMC proliferation in response to arterial cyclic stretch.Therefore,miRNA-33 may be a potential therapeutic target in autologous vein grafted surgery,and locally overexpression of miR-33 may attenuates neointimal hyperplasia of grafted human saphenous vein [3].The unpublished data revealed that 15%cyclic stretch also significantly elevated the expression of miRNA-124-3p which bound to the 3’UTR of Lmna mRNA,and then negatively regulated protein expression of lamin A/C which is the important skeletal proteins in nucleus.In addition to primary intracellular locations of miRNAs,our recent study showed that miRNAs can be secreted and protected extracellularly via inclusion into membrane-derived vesicles including microparticles.Microparticles are extracellular vesicles ranging from 0.1 to 1μm in size and have been shown to deliver various bioactive molecules,i.e.,chemokines,enzymes and miRNAs,to recipient cells.Increasing evidence shows that microparticles play a pivotal role in many pathological processes,such as cancer,inflammatory diseases and cardiovascular disease.Our present study showed that platelet-derived microparticles(PMPs),which are released by active platelets,are important vehicles for communication and play crucial roles in inducing abnormal EC proliferation in hypertension.In briefly,EC proliferation was increased in renal hypertensive rats established by abdominal aortic coarctation compared to control rats and that elevated thrombin in plasma promoted platelet activation,which may induce the release of PMPs.miRNA array and qPCR revealed a higher level of miRNA-142-3p in platelets and PMPs.In vitro,PMPs delivered miRNA-142-3p into ECs and enhanced EC proliferation via Bcl-2-associated transcription factor 1(BCLAF1)and its downstream genes.These results indicated that PMPs deliver miRNA-142-3p from activated platelets into ECs and that miRNA-142-3p may play important roles in EC dysfunction under hypertensive conditions and might be a novel therapeutic target for maintaining EC homeostasis in hypertension[1].These results provide possible mechanisms by which non-coding RNAs regulate cellular functions under different mechanical stresses,and suggest a novel potential therapeutic approach for vascular remodeling.The further studies on noncoding RNAs may provide new insight into understanding the mechanism of vascular remodeling in different various cardiovascular disorders,and may provide novel targets for the maintenance of vascular homeostasis.
文摘以高耐热性玉米品种郑单958、低耐热性玉米品种先玉335为试验材料,以正常生长条件为对照(CK),利用半自动伸缩高温棚进行花期高温胁迫(HT)处理,通过circRNA高通量测序筛选高温胁迫下不同玉米品种花粉中差异表达的环状RNA(circRNA),对其来源基因进行GO和KEGG富集分析,并筛选具有miRNA结合位点的差异表达circRNA,预测其下游目的基因,分析玉米花粉中响应高温胁迫的潜在circRNA-miRNA-mRNA共表达调控网络,从多层面解析玉米花粉中调控高温胁迫的分子作用机制,为提高玉米品种的耐热性提供理论依据。结果表明,在郑单958、先玉335不同样本中共鉴定出1 843个不同的circRNA,它们在玉米染色体中的分布不同。每个circRNA所包含的外显子数目也不相同,其中,大多数(624个)circRNA只含有1个外显子。在郑单958花粉中共鉴定出1 563个circRNA,其中,CK958-1、CK958-2、CK958-3中分别鉴定出305、213、356个circRNA,HT958-1、HT958-2、HT958-3中分别鉴定出222、242、225个circRNA。在先玉335花粉中共鉴定出1 423个circRNA,其中,CK335-1、CK335-2、CK335-3中分别鉴定出272、188、229个circRNA,HT335-1、HT335-2、HT335-3中分别鉴定出259、237、238个circRNA。不同样本中占比最高的均为外显子circRNA。circRNA与其来源基因不是一一对应的关系,有748个circRNA来源基因通过反向剪接机制只形成1个circRNA,156个circRNA来源基因通过反向剪接机制各自形成2个circRNA。在郑单958高温胁迫花粉与对照花粉对比组(HT958 vs CK958)中共筛选到9个差异表达circRNA,其中2个circRNA呈上调表达,其来源基因显著富集到焦磷酸酶活性、核苷酸磷酸代谢过程、糖基磷脂酰肌醇(GPI)锚定代谢过程等17个GO条目,显著富集到GPI锚定生物合成、代谢途径等KEGG通路。在先玉335高温胁迫花粉与对照花粉对比组(HT335 vs CK335)中共筛选到1个差异表达circRNA,其来源基因没有显著富集到任何GO条目、KEGG通路。在郑单958高温胁迫花粉与先玉335高温胁迫花粉对比组(HT958 vs HT335)中共筛选到17个差异表达circRNA,其中6个circRNA呈上调表达,其来源基因显著富集到内质网系统、高尔基相关囊泡膜、膜蛋白水解等16个GO条目中,没有显著富集到任何KEGG代谢通路。5个circRNA具有miRNA结合位点,可以作为海绵岛吸附miRNA间接调控下游靶标基因的表达,构建了包括5个circRNA、5个不同家族miRNA、2个mRNA在内的circRNA-miRNA-mRNA共表达调控网络。筛选到了54个circRNA包含内部核糖体进入位点(IRES),可以翻译表达多肽或者蛋白质直接作用于靶标基因。
