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
目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X rec...目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。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。
文摘目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。
