Objectives Autologous blood outgrowth endothelial cells(BOECs)have been proposed to induce therapeutic angiogenesis for treating cardiovascular diseases(CVDs).The aim of the present study was to investigate the prolif...Objectives Autologous blood outgrowth endothelial cells(BOECs)have been proposed to induce therapeutic angiogenesis for treating cardiovascular diseases(CVDs).The aim of the present study was to investigate the proliferative potential and angiogenic characteristics of BOECs among middle-aged and older adults,the population particularly susceptible to CVDs.Methods BOECs were isolated from 48 peripheral blood samples of subjects aged 56±4 years.The cells were then distinguished based on their proliferative abilities,and their phenotype,tube formation capacity,and migratory activity were compared using immunofluorescence staining,flow cytometry,tube formation assay,and wound healing assay,respectively.Correlations between demographic,clinical,and dietary parameters with the number of BOECs were also assessed.Results A total of 132 BOEC colonies with different proliferative potentials were obtained,including colonies lost proliferative ability before passage 3(named LPA),stopped proliferating during passage 3–8(HPA(3–8)),and proliferated after passage 8(HPA(>8)).LPA cells appeared later and displayed abnormal morphology,while HPA(3–8)cells exhibited alterations in von Willebrand factor morphology and lower KDR expression.HPA(>8)cells obtained higher branching intervals and individual cell migration velocity compared with those of HPA(3–8)cells.Correlation analysis showed that the number of both LPA and HPA colonies were positively associated with several CVD risk factors.Additionally,the number of LPA colonies was positively associated with servings of meats and alternatives,fruits,fruits and vegetables,as well as the protein intake.Conclusions Our findings provide evidence that the middle-aged and older populations possess BOECs with different proliferative and angiogenic potentials,exhibiting distinctions in cell morphology,appearance dates,VWF morphology,and KDR expression.Strikingly,a higher number of BOECs is likely associated with an increased risk of CVDs,while the number of BOECs with low proliferative ability may be regulated by diet.展开更多
Background:Pancreatic cancer(PC)is a highly deadly malignancy with few effective therapies.We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6(SNHG6)plays in PC cells by targeting far ...Background:Pancreatic cancer(PC)is a highly deadly malignancy with few effective therapies.We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6(SNHG6)plays in PC cells by targeting far upstream element binding protein 1(FUBP1)via microRNA-26a-5p(miR-26a-5p).Methods::SNHG6 expression was predicted by bioinformatics,followed by verification via reverse transcription quantitative polymerase chain reaction.Then,the interactions among SNHG6,miR-26a-5p,and FUBP1 were detected through online software analysis,dual luciferase reporter assay and RNA pull-down.After that,cells were treated with different small interfering RNAs and/or mimic to determine the interactions among SNHG6,miR-26a-5p,and FUBP1 and their roles in PC cells.Finally,the role of SNHG6 in tumor growth in vivo was evaluated by measuring the growth and weight of transplanted tumors in nude mice.A t-test,one-way and two-way analysis of variance were used for data analysis.Results::Compared with that in normal tissues,SNHG6 was highly expressed in PC tissues(1.00±0.05 vs.1.56±0.06,t=16.03,P<0.001).Compared with that in human pancreatic duct epithelial cells(HPDE6-C7),SNHG6 showed the highest expression in PANC-1 cells(1.00±0.06 vs.3.87±0.13,t=34.72,P<0.001)and the lowest expression in human pancreatic cancer cells(MIAPaCa-2)(1.00±0.06 vs.1.41±0.07,t=7.70,P=0.0015).Compared with the levels in the si-negative control group,SNHG6(0.97±0.05 vs.0.21±0.06,t=16.85,P<0.001),N-cadherin(0.74±0.05 vs.0.41±0.04,t=8.93,P<0.001),Vimentin(0.55±0.04 vs.0.25±0.03,t=10.39,P<0.001),andβ-catenin(0.62±0.05 vs.0.32±0.03,t=8.91,P<0.001)were decreased,while E-cadherin(0.65±0.06 vs.1.36±0.07,t=13.34,P<0.001)was increased after SNHG6 knockdown or miR-26a-5p overexpression,accompanied by inhibited cell proliferation,migration,and invasion.SNHG6 overexpression exerted the opposite effects.SNHG6 upregulated FUBP1 expression by sponging miR-26a-5p.Silencing SNHG6 blocked the growth of PC in vivo.Conclusion::Silencing SNHG6 might ameliorate PC through inhibition of FUBP1 by sponging miR-26a-5p,thus providing further supporting evidence for its use in PC treatment.展开更多
基金sponsored by Shanghai Pujiang Program(23PJ1409600)Intergovernmental International Science and Technology Innovation Cooperation Key Project of Chinese National Key R&D Program(2024YFE0102600)Singapore Ministry of Education Academic Research Fund Tier 1(R160-000-A26–114).
文摘Objectives Autologous blood outgrowth endothelial cells(BOECs)have been proposed to induce therapeutic angiogenesis for treating cardiovascular diseases(CVDs).The aim of the present study was to investigate the proliferative potential and angiogenic characteristics of BOECs among middle-aged and older adults,the population particularly susceptible to CVDs.Methods BOECs were isolated from 48 peripheral blood samples of subjects aged 56±4 years.The cells were then distinguished based on their proliferative abilities,and their phenotype,tube formation capacity,and migratory activity were compared using immunofluorescence staining,flow cytometry,tube formation assay,and wound healing assay,respectively.Correlations between demographic,clinical,and dietary parameters with the number of BOECs were also assessed.Results A total of 132 BOEC colonies with different proliferative potentials were obtained,including colonies lost proliferative ability before passage 3(named LPA),stopped proliferating during passage 3–8(HPA(3–8)),and proliferated after passage 8(HPA(>8)).LPA cells appeared later and displayed abnormal morphology,while HPA(3–8)cells exhibited alterations in von Willebrand factor morphology and lower KDR expression.HPA(>8)cells obtained higher branching intervals and individual cell migration velocity compared with those of HPA(3–8)cells.Correlation analysis showed that the number of both LPA and HPA colonies were positively associated with several CVD risk factors.Additionally,the number of LPA colonies was positively associated with servings of meats and alternatives,fruits,fruits and vegetables,as well as the protein intake.Conclusions Our findings provide evidence that the middle-aged and older populations possess BOECs with different proliferative and angiogenic potentials,exhibiting distinctions in cell morphology,appearance dates,VWF morphology,and KDR expression.Strikingly,a higher number of BOECs is likely associated with an increased risk of CVDs,while the number of BOECs with low proliferative ability may be regulated by diet.
基金This study was supported by a grant from the Shanghai University of Medicine&Health Sciences Seed Foundation(No.SFP-18-22-15-002).
文摘Background:Pancreatic cancer(PC)is a highly deadly malignancy with few effective therapies.We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6(SNHG6)plays in PC cells by targeting far upstream element binding protein 1(FUBP1)via microRNA-26a-5p(miR-26a-5p).Methods::SNHG6 expression was predicted by bioinformatics,followed by verification via reverse transcription quantitative polymerase chain reaction.Then,the interactions among SNHG6,miR-26a-5p,and FUBP1 were detected through online software analysis,dual luciferase reporter assay and RNA pull-down.After that,cells were treated with different small interfering RNAs and/or mimic to determine the interactions among SNHG6,miR-26a-5p,and FUBP1 and their roles in PC cells.Finally,the role of SNHG6 in tumor growth in vivo was evaluated by measuring the growth and weight of transplanted tumors in nude mice.A t-test,one-way and two-way analysis of variance were used for data analysis.Results::Compared with that in normal tissues,SNHG6 was highly expressed in PC tissues(1.00±0.05 vs.1.56±0.06,t=16.03,P<0.001).Compared with that in human pancreatic duct epithelial cells(HPDE6-C7),SNHG6 showed the highest expression in PANC-1 cells(1.00±0.06 vs.3.87±0.13,t=34.72,P<0.001)and the lowest expression in human pancreatic cancer cells(MIAPaCa-2)(1.00±0.06 vs.1.41±0.07,t=7.70,P=0.0015).Compared with the levels in the si-negative control group,SNHG6(0.97±0.05 vs.0.21±0.06,t=16.85,P<0.001),N-cadherin(0.74±0.05 vs.0.41±0.04,t=8.93,P<0.001),Vimentin(0.55±0.04 vs.0.25±0.03,t=10.39,P<0.001),andβ-catenin(0.62±0.05 vs.0.32±0.03,t=8.91,P<0.001)were decreased,while E-cadherin(0.65±0.06 vs.1.36±0.07,t=13.34,P<0.001)was increased after SNHG6 knockdown or miR-26a-5p overexpression,accompanied by inhibited cell proliferation,migration,and invasion.SNHG6 overexpression exerted the opposite effects.SNHG6 upregulated FUBP1 expression by sponging miR-26a-5p.Silencing SNHG6 blocked the growth of PC in vivo.Conclusion::Silencing SNHG6 might ameliorate PC through inhibition of FUBP1 by sponging miR-26a-5p,thus providing further supporting evidence for its use in PC treatment.
