Background Hypoxic stimuli induce follicular atresia by regulating granulosa cell(GC)apoptosis.Notably,mature follicles can still develop and ovulate under hypoxic conditions,highlighting the importance of the hypoxic...Background Hypoxic stimuli induce follicular atresia by regulating granulosa cell(GC)apoptosis.Notably,mature follicles can still develop and ovulate under hypoxic conditions,highlighting the importance of the hypoxic adaptation in ovarian follicular selection.To date,the role and mechanism of hypoxia-inducible factor 1 subunit alpha(HIF1A)-mediated hypoxic responses in follicular atresia are unclear.This study aimed to investigate whether and how HIF1A regulates follicular atresia via the modulation of O-linked N-acetylglucosamine(O-GlcNAc)protein modification(O-GlcNAcylation).Results Our findings revealed that HIF1A was highly expressed in pig ovaries.Compared with that in healthy follicles,its expression was significantly downregulated in atretic follicles.Under hypoxic conditions,pharmacological inhibition or siRNA-mediated knockdown of HIF1A increased porcine GC apoptosis.Mechanistically,HIF1A knockdown Suppressed O-GlcNAc transferase degradation,leading to increased global O-GlcNAcylation.Using 4D labelfree quantitative proteomics,we identified 53 O-GlcNAcylated proteins.Importantly,O-GlcNAcylation stabilized vascular endothelial zinc finger 1(VEZF1),and HIF1A knockdown upregulated VEZF1 protein levels by promoting O-GlcNAcylation.The HIF1A-VEZF1 axis modulates forkhead box O1(FOXO1)expression by regulating endothelin-1.As a transcription factor,FOXO1 directly binds to the Bcl-2 associated X(BAX)promoter,activating its transcription and ultimately inducing porcine GC apoptosis and follicular atresia.Conclusion Overall,our study elucidates a novel molecular mechanism by which HIF1A deficiency modulates follicular atresia through O-GlcNAcylation-mediated VEZF1 expression.These results not only clarify the molecular mechanism of ovarian follicular development under hypoxic conditions but also offer potential targets for improving follicular selection efficiency in pig breeding.展开更多
Objective:Several reports have proposed that lnc RNAs,as potential biomarkers,participate in the progression and growth of malignant tumors.HIF1 A-AS2 is a novel lnc RNA and potential biomarker,involved in the genesis...Objective:Several reports have proposed that lnc RNAs,as potential biomarkers,participate in the progression and growth of malignant tumors.HIF1 A-AS2 is a novel lnc RNA and potential biomarker,involved in the genesis and development of carcinomas.However,the molecular mechanism of HIF1 A-AS2 in renal carcinoma is unclear.Methods:The relative expression levels of HIF1 A-AS2 and miR-30 a-5 p were detected using RT-qPCR in renal carcinoma tissues and cell lines.Using loss-of-function and overexpression,the biological effects of HIF1 A-AS2 and miR-30 a-5 p in kidney carcinoma progression were characterized.Dual luciferase reporter gene analysis and Western blot were used to detect the potential mechanism of HIF1 A-AS2 in renal carcinomas.Results:HIF1 A-AS2 was upregulated in kidney carcinoma tissues when compared with para-carcinoma tissues(P<0.05).In addition,tumor size,tumor node mestastasis stage and differentiation were identified as being closely associated with HIF1 A-AS2 expression(P<0.05).Knockdown or overexpression of HIF1 A-AS2 either restrained or promoted the malignant phenotype and WNT/β-catenin signaling in renal carcinoma cells(P<0.05).Mi R-30 a-5 p was downregulated in renal cancers and partially reversed HIF1 A-AS2 functions in malignant renal tumor cells.HIF1 A-AS2 acted as a micro RNA sponge that actively regulated the relative expression of SOX4 in sponging miR-30 a-5 p and subsequently increased the malignant phenotypes of renal carcinomas.HIF1 A-AS2 showed a carcinogenic effect and miR-30 a-5 p acted as an antagonist of the anti-oncogene effects in the pathogenesis of renal carcinomas.Conclusions:The HIF1 A-AS2-miR-30 a-5 p-SOX4 axis was associated with the malignant progression and development of renal carcinoma.The relative expression of HIF1 A-AS2 was negatively correlated with the expression of miR-30 a-5 p,and was closely correlated with SOX4 mRNA levels in renal cancers.展开更多
基金supported by the National Natural Science Foundation of China(32402710)National Postdoctoral Researchers Funding Program(GZC20232209)+1 种基金Higher Education Institutions Basic Science(Natural Science)Research General Program of Jiangsu Province(24KJB230005)Priority Academic Program Development of Jiangsu Higher Education Institution.
文摘Background Hypoxic stimuli induce follicular atresia by regulating granulosa cell(GC)apoptosis.Notably,mature follicles can still develop and ovulate under hypoxic conditions,highlighting the importance of the hypoxic adaptation in ovarian follicular selection.To date,the role and mechanism of hypoxia-inducible factor 1 subunit alpha(HIF1A)-mediated hypoxic responses in follicular atresia are unclear.This study aimed to investigate whether and how HIF1A regulates follicular atresia via the modulation of O-linked N-acetylglucosamine(O-GlcNAc)protein modification(O-GlcNAcylation).Results Our findings revealed that HIF1A was highly expressed in pig ovaries.Compared with that in healthy follicles,its expression was significantly downregulated in atretic follicles.Under hypoxic conditions,pharmacological inhibition or siRNA-mediated knockdown of HIF1A increased porcine GC apoptosis.Mechanistically,HIF1A knockdown Suppressed O-GlcNAc transferase degradation,leading to increased global O-GlcNAcylation.Using 4D labelfree quantitative proteomics,we identified 53 O-GlcNAcylated proteins.Importantly,O-GlcNAcylation stabilized vascular endothelial zinc finger 1(VEZF1),and HIF1A knockdown upregulated VEZF1 protein levels by promoting O-GlcNAcylation.The HIF1A-VEZF1 axis modulates forkhead box O1(FOXO1)expression by regulating endothelin-1.As a transcription factor,FOXO1 directly binds to the Bcl-2 associated X(BAX)promoter,activating its transcription and ultimately inducing porcine GC apoptosis and follicular atresia.Conclusion Overall,our study elucidates a novel molecular mechanism by which HIF1A deficiency modulates follicular atresia through O-GlcNAcylation-mediated VEZF1 expression.These results not only clarify the molecular mechanism of ovarian follicular development under hypoxic conditions but also offer potential targets for improving follicular selection efficiency in pig breeding.
基金supported by grants from the National Natural Science Foundations of China(Grant Nos.81702511,81472401,81772708,and 2016YFA0201204)the Jiangsu Provincial Key Medical Discipline(Grant No.ZDXKA2016012)+1 种基金the Clinical Medicine Center of Suzhou(Grant No.SZZXJ201501)programs for Recruitment of Clinical Medical Top Team of Suzhou。
文摘Objective:Several reports have proposed that lnc RNAs,as potential biomarkers,participate in the progression and growth of malignant tumors.HIF1 A-AS2 is a novel lnc RNA and potential biomarker,involved in the genesis and development of carcinomas.However,the molecular mechanism of HIF1 A-AS2 in renal carcinoma is unclear.Methods:The relative expression levels of HIF1 A-AS2 and miR-30 a-5 p were detected using RT-qPCR in renal carcinoma tissues and cell lines.Using loss-of-function and overexpression,the biological effects of HIF1 A-AS2 and miR-30 a-5 p in kidney carcinoma progression were characterized.Dual luciferase reporter gene analysis and Western blot were used to detect the potential mechanism of HIF1 A-AS2 in renal carcinomas.Results:HIF1 A-AS2 was upregulated in kidney carcinoma tissues when compared with para-carcinoma tissues(P<0.05).In addition,tumor size,tumor node mestastasis stage and differentiation were identified as being closely associated with HIF1 A-AS2 expression(P<0.05).Knockdown or overexpression of HIF1 A-AS2 either restrained or promoted the malignant phenotype and WNT/β-catenin signaling in renal carcinoma cells(P<0.05).Mi R-30 a-5 p was downregulated in renal cancers and partially reversed HIF1 A-AS2 functions in malignant renal tumor cells.HIF1 A-AS2 acted as a micro RNA sponge that actively regulated the relative expression of SOX4 in sponging miR-30 a-5 p and subsequently increased the malignant phenotypes of renal carcinomas.HIF1 A-AS2 showed a carcinogenic effect and miR-30 a-5 p acted as an antagonist of the anti-oncogene effects in the pathogenesis of renal carcinomas.Conclusions:The HIF1 A-AS2-miR-30 a-5 p-SOX4 axis was associated with the malignant progression and development of renal carcinoma.The relative expression of HIF1 A-AS2 was negatively correlated with the expression of miR-30 a-5 p,and was closely correlated with SOX4 mRNA levels in renal cancers.
