Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-t...Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.展开更多
Background Within the follicular fluid,extracellular vesicles(EVs)guide oocyte growth through their cargo microRNAs(miRNAs).Here,we inv estigated the role of EVs and their cargo miRNAs by linking the miRNAs found in E...Background Within the follicular fluid,extracellular vesicles(EVs)guide oocyte growth through their cargo microRNAs(miRNAs).Here,we inv estigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs,derived from the fluid of an individual follicle,to the ability of its oocyte to become a blastocyst(competent)or not(non-competent).Methods Bovine antral follicles were dissected,categorized as small(2-4 mm)or large(5-8 mm)and the corresponding oocytes were subjected to individual maturation,fertilization and embryo culture to the blastocyst stage.Follicular fluid was pooled in 4 groups(4 replicates)based on follicle size and competence of the corresponding oocyte to produce a blastocyst.Follicular fluid-derived EVs were isolated",characterized,and subjected to miRNAsequencing(Illumina Miseq)to assess differential expression(DE)in the 4 groups.Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation(IVM).Results We identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered.Within the large and small follicles,46 DE miRNAs were driving blastocyst formation in each group.Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs.Cell regulation,cell differentiation,cell cycle,and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes.We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes.Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development.However,blastocyst quality,as evidenced by higher cell numbers,was significantly improved following oocyte IVM in the presence of miR-34c mimics,while miR-34c inhibitors resulted in the opposite effect.Conclusion This study demonstrates the regulatory effect of miRNAs from follicular fliived EVs on oocyte competence acquisitividing a further basis for understanding the significance of miRNAs in oocyte maturation and embryonic development Up-regulation of miR-34c in EVs from follicular fluid containing competent oocytes and the positive impact of miR-34c mimics added during IVM on the resulting blastocysts indicate its pivotal role in oocyte competence.展开更多
基金supported by the Medical Science and Technology Research Foundation of Guangdong Province(No.A2020559).
文摘Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.
基金supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 860960by Bijzonder Onderzoeksfonds GOA(Geconcerteerde onderzoeksacties)2018000504(GOA030-18 BOF)supported by the Research Foundation Flanders(FWO)(grant numbers:1228821N and 12A2H24N)。
文摘Background Within the follicular fluid,extracellular vesicles(EVs)guide oocyte growth through their cargo microRNAs(miRNAs).Here,we inv estigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs,derived from the fluid of an individual follicle,to the ability of its oocyte to become a blastocyst(competent)or not(non-competent).Methods Bovine antral follicles were dissected,categorized as small(2-4 mm)or large(5-8 mm)and the corresponding oocytes were subjected to individual maturation,fertilization and embryo culture to the blastocyst stage.Follicular fluid was pooled in 4 groups(4 replicates)based on follicle size and competence of the corresponding oocyte to produce a blastocyst.Follicular fluid-derived EVs were isolated",characterized,and subjected to miRNAsequencing(Illumina Miseq)to assess differential expression(DE)in the 4 groups.Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation(IVM).Results We identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered.Within the large and small follicles,46 DE miRNAs were driving blastocyst formation in each group.Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs.Cell regulation,cell differentiation,cell cycle,and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes.We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes.Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development.However,blastocyst quality,as evidenced by higher cell numbers,was significantly improved following oocyte IVM in the presence of miR-34c mimics,while miR-34c inhibitors resulted in the opposite effect.Conclusion This study demonstrates the regulatory effect of miRNAs from follicular fliived EVs on oocyte competence acquisitividing a further basis for understanding the significance of miRNAs in oocyte maturation and embryonic development Up-regulation of miR-34c in EVs from follicular fluid containing competent oocytes and the positive impact of miR-34c mimics added during IVM on the resulting blastocysts indicate its pivotal role in oocyte competence.
