OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like g...OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.展开更多
目的探讨孕妇血清胰岛素样生长因子1(insulin like growth factor 1,IGF-1)、IGF-2、IGF结合蛋白3(IGF binding protein 3,IGFBP-3)与正常胎儿生长的关系。方法选择2010年1月至2011年5月于上海市浦东新区人民医院产前检查并分娩正常体...目的探讨孕妇血清胰岛素样生长因子1(insulin like growth factor 1,IGF-1)、IGF-2、IGF结合蛋白3(IGF binding protein 3,IGFBP-3)与正常胎儿生长的关系。方法选择2010年1月至2011年5月于上海市浦东新区人民医院产前检查并分娩正常体重儿的初产妇66例,分为妊娠16～18周、妊娠26～28周、妊娠37～40周3个阶段进行纵向观察,放射免疫法测定孕妇各阶段血清中IGF-1、IGF-2、IGFBP-3水平并进行对比分析。结果孕期母血IGF-1水平随着孕周增加明显上升,其中IGF-1水平在妊娠37～40周高于妊娠26～28周,妊娠26～28周高于妊娠16～18周,差异均有显著统计学意义(P均<0.01)。孕期母血IGF-2水平随孕周增加无明显改变,妊娠3阶段差异无统计学意义(P>0.05)。母血IGFBP-3水平妊娠37～40周高于妊娠26～28周及妊娠16～18周期,差异有统计学意义(P<0.05),而妊娠26～28周与妊娠16～18周无显著差异。妊娠16～18周、26～28周和37～40周3阶段母血IGF-1、IGF-2、IGFBP-3水平与正常新生儿出生体重无显著相关性。结论孕妇血清IGF-1、IGFBP-3水平与正常胎儿生长密切相关,IGF-1可作为临床评价不同阶段正常胎儿生长的指标,而IGFBP-3更多地反映了妊娠中晚期正常胎儿的生长。展开更多
目的明确外源性胰岛素样生长因子1(IGF-1)多肽对体外培养的人牙周膜干细胞(hPDLSC)增殖及分化的影响。方法有限稀释法分离培养hPDLSC,流式细胞术检测细胞表面标志物的表达。结晶紫染色观察细胞集落形成。采用成骨及成脂诱导液诱导hPDLS...目的明确外源性胰岛素样生长因子1(IGF-1)多肽对体外培养的人牙周膜干细胞(hPDLSC)增殖及分化的影响。方法有限稀释法分离培养hPDLSC,流式细胞术检测细胞表面标志物的表达。结晶紫染色观察细胞集落形成。采用成骨及成脂诱导液诱导hPDLSC分化。采用CCK-8法分析(1、10、30、50、100)ng/m L IGF-1处理对hPDLSC增殖活性的影响。实时定量PCR检测IGF-1处理的hPDLSC中1型胶原蛋白α1(Col1α1)、3型胶原蛋白α1(Col3α1)、抗Runt相关转录因子2(Runx2)、骨涎蛋白(BSP)、骨桥蛋白(OPN)、牙骨质蛋白1(CEMP1)、scleraxis(SCXA)m RNA的表达。Western blot法检测Col1、Col3、Runx2、CEMP1及SCXA蛋白表达的水平。结果有限稀释法分离培养的hPDLSC高表达干细胞表面标志物CD105(95.4%)和CD90(96.1%),而低表达造血细胞标志物CD34(1.58%)、CD45(1.74%)。hPDLSC可形成集落并具有向成骨和成脂分化能力。IGF-1可提高hPDLSC的增殖活性,并呈剂量依赖效应。与对照组相比,IGF-1处理的hPDLSC中Runx2、BSP、OPN及CEMP1 m RNA的表达水平无明显变化,而Col1α1、Col3α1、SCXA显著增加。并且IGF-1处理后hPDLSC中Col1、Col3及SCXA的蛋白含量增加。结论 IGF-1可促进hPDLSC增殖和胶原蛋白合成,加速其向牙周膜成纤维细胞分化。展开更多
目的探讨电针听宫穴和翳风穴对豚鼠听皮层葡萄糖代谢和胰岛素样生长因子-1(insulin like growth factor-1,IGF-1)表达的影响。方法将成年杂色豚鼠18只随机分为3组,电针耳穴组7只,同时电针双侧听宫和翳风穴,持续15分钟,每日1次,共7天;电...目的探讨电针听宫穴和翳风穴对豚鼠听皮层葡萄糖代谢和胰岛素样生长因子-1(insulin like growth factor-1,IGF-1)表达的影响。方法将成年杂色豚鼠18只随机分为3组,电针耳穴组7只,同时电针双侧听宫和翳风穴,持续15分钟,每日1次,共7天;电针非穴位组6只,电针听宫穴靠眼侧方5mm处和翳风穴正下方5mm处,时间同电针耳穴组;对照组5只不作任何处理,常规饲养。各组完成电针后进行PET/CT扫描,获得CT、PET及两者融合图像,观察豚鼠听皮层摄取显像剂18F-脱氧葡萄糖的情况,以听皮层和小脑最大标准摄入值(standard uptake value,SUV)的比值为观察指标;采用实时荧光定量PCR技术检测豚鼠听皮层IGF-1mRNA表达。结果对照组、电针耳穴组、电针非耳穴组听皮层SUV最大值与小脑SUV最大值的比值分别为0.75±0.06、0.84±0.05和0.71±0.06,电针耳穴组高于对照组和电针非耳穴组,(分别为P=0.042,P=0.009);对照组、电针耳穴组、电针非耳穴组听皮层局部IGF-1mRNA相对表达量分别为1.34±0.24、2.03±0.36、0.92±0.23,电针耳穴组高于对照组和电针非耳穴组(分别为P=0.002,P<0.001)。结论电针针刺听宫和翳风穴可增加豚鼠听皮层局部IGF-1的表达,可能通过提高葡萄糖代谢水平促进豚鼠听皮层的功能。展开更多
基金supported by National Natural Science Foundation of China(81502123 and81330081)Natural Science Foundation of Anhui Province(1308085QH130)Anhui Province Nature Science Foundation in University(KJ2014A119)
文摘OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.
文摘目的探讨孕妇血清胰岛素样生长因子1(insulin like growth factor 1,IGF-1)、IGF-2、IGF结合蛋白3(IGF binding protein 3,IGFBP-3)与正常胎儿生长的关系。方法选择2010年1月至2011年5月于上海市浦东新区人民医院产前检查并分娩正常体重儿的初产妇66例,分为妊娠16～18周、妊娠26～28周、妊娠37～40周3个阶段进行纵向观察,放射免疫法测定孕妇各阶段血清中IGF-1、IGF-2、IGFBP-3水平并进行对比分析。结果孕期母血IGF-1水平随着孕周增加明显上升,其中IGF-1水平在妊娠37～40周高于妊娠26～28周,妊娠26～28周高于妊娠16～18周,差异均有显著统计学意义(P均<0.01)。孕期母血IGF-2水平随孕周增加无明显改变,妊娠3阶段差异无统计学意义(P>0.05)。母血IGFBP-3水平妊娠37～40周高于妊娠26～28周及妊娠16～18周期,差异有统计学意义(P<0.05),而妊娠26～28周与妊娠16～18周无显著差异。妊娠16～18周、26～28周和37～40周3阶段母血IGF-1、IGF-2、IGFBP-3水平与正常新生儿出生体重无显著相关性。结论孕妇血清IGF-1、IGFBP-3水平与正常胎儿生长密切相关,IGF-1可作为临床评价不同阶段正常胎儿生长的指标,而IGFBP-3更多地反映了妊娠中晚期正常胎儿的生长。
文摘目的明确外源性胰岛素样生长因子1(IGF-1)多肽对体外培养的人牙周膜干细胞(hPDLSC)增殖及分化的影响。方法有限稀释法分离培养hPDLSC,流式细胞术检测细胞表面标志物的表达。结晶紫染色观察细胞集落形成。采用成骨及成脂诱导液诱导hPDLSC分化。采用CCK-8法分析(1、10、30、50、100)ng/m L IGF-1处理对hPDLSC增殖活性的影响。实时定量PCR检测IGF-1处理的hPDLSC中1型胶原蛋白α1(Col1α1)、3型胶原蛋白α1(Col3α1)、抗Runt相关转录因子2(Runx2)、骨涎蛋白(BSP)、骨桥蛋白(OPN)、牙骨质蛋白1(CEMP1)、scleraxis(SCXA)m RNA的表达。Western blot法检测Col1、Col3、Runx2、CEMP1及SCXA蛋白表达的水平。结果有限稀释法分离培养的hPDLSC高表达干细胞表面标志物CD105(95.4%)和CD90(96.1%),而低表达造血细胞标志物CD34(1.58%)、CD45(1.74%)。hPDLSC可形成集落并具有向成骨和成脂分化能力。IGF-1可提高hPDLSC的增殖活性,并呈剂量依赖效应。与对照组相比,IGF-1处理的hPDLSC中Runx2、BSP、OPN及CEMP1 m RNA的表达水平无明显变化,而Col1α1、Col3α1、SCXA显著增加。并且IGF-1处理后hPDLSC中Col1、Col3及SCXA的蛋白含量增加。结论 IGF-1可促进hPDLSC增殖和胶原蛋白合成,加速其向牙周膜成纤维细胞分化。
