The myeloid zinc finger gene-1 (MZF-1) encodes a putative transcription factor whose expression has been implicated in myeloid differentiation. To study the role of the nMZF-1 in myploid differentiation,we characteriz...The myeloid zinc finger gene-1 (MZF-1) encodes a putative transcription factor whose expression has been implicated in myeloid differentiation. To study the role of the nMZF-1 in myploid differentiation,we characterized MZF-1 protein expr.ession,cellular localization,and phosphorylation in leukemia cell lines and leukemia cells.MZF1 protein expression was found only in myeloid cells. In proliferating HL-60 cells,MZF-1 was localized to the nucleus with some cytoplasmic distribution; however,upon retinoic acid (RA)induced granulocytic differentiation, MZF-1 became restricted to the nucleus.In32 PO4-la labelled HL-60 cell, MZF-1 was immunoprecipitated as a phosphoprotein doublet of 53￣54kDa. MZF-1 phosphorylation increased after acute stimulation of HL-60 with granulocytemacrophage colony stimulating factor (GM-CSF), interleukin-3(IL-3),phorbol ester,and serum.Chronic GM-CSf treatment of HL-60 cells potentiating granulocytic differentiation sustained the hyperphosphorylated state of MZF-1,whereas chronic treatment with TPA leading to monocytic-macrophage differentiation was accompanied by the disappearance of the 53 kDa MZF-1 phosphoprotein and the appearance of cross-reactive 69 and 105kDa phosphoprotein species. K562 human myeloblastic cells which are resistant to granulocytic differentiation express both the 53 kDa MZF-1 protein and the cross reactive 69 and 105 kDa proteins,but the 53 kDa MZF-1 protein is not detectable phosphorylated under any experimental conditions. Acute promyelocytic leukemic cells exhibited the 53kDa phosphoprotein,whereas monocytic leukemia cells expressed only the 69 and 105 kDa MZF-1 related phosphoproteins. The studies demonstrate that MZF-1 is a nuclear protein whose phosphorylation is associated with the granulocytic commitment of myeloid cells.展开更多
AIM： To investigate the promoter region methylation status of retinoblastoma protein-interacting zinc finger gene 1 （RIZ1） in the human esophageal squamous cell carcinoma （ESCC） cell lines and tissues and verify ...AIM： To investigate the promoter region methylation status of retinoblastoma protein-interacting zinc finger gene 1 （RIZ1） in the human esophageal squamous cell carcinoma （ESCC） cell lines and tissues and verify the relationship between methylation of RIZ1 and oncogen- esis, tumor progression and metastasis etc of ESCC. METHODS： Methylation-specific polymerase chain reac- tion （MSP） was used to investigate the promoter region methylation status of RIZ1 in 6 ESCC cell lines. One cell line where RIZ1 promoter region methylation was de- tected was selected for the next study, where the cell line was treated with 5-aza-CdR. Real-time polymerase chain reaction was used to investigate its influence on the transcription of RIZ1. Experiments using frozenpathological specimens from 47 ESCC patients were performed using the same MSP methodology. RESULTS： Promoter methylation of RIZ1 gene was detected in TEl3, CaEs17 and EC109 cell lines and the cell line TEl3 was chosen for further study. The expression of RIZl mRNA in TE-13 was up-regulated after treatment with 5-aza-CdR. The rate of methyla- tion in carcinomas tissues was significantly higher than those in matched neighboring normal and distal ending normal tissue, and the deviation of data was statisti- cally significant （2,2 = 24.136, P 〈 0.01）. Analysis of the gender, age familial history, tumour deviation, tumour saturation, lymph gland displacement and clinical stag- ing of 47 samples from ESCC patients showed that the fluctuation of data was not statistically significant. CONCLUSION： Promoter methylation may play an im- portant role in the epigenetic silencing of RIZ1 gene expression in human ESCC. RIZ1 is considered to be a potential tumor suppressor gene and may be a biologi- cal parameter for testing early stage human ESCC.展开更多
AIM:To investigate the effect of retinoblastoma protein-interacting zinc finger gene 1(RIZ1)upregulation in gene expression profile and oncogenicity of human esophageal squamous cell carcinoma(ESCC)cell line TE13.METH...AIM:To investigate the effect of retinoblastoma protein-interacting zinc finger gene 1(RIZ1)upregulation in gene expression profile and oncogenicity of human esophageal squamous cell carcinoma(ESCC)cell line TE13.METHODS:TE13 cells were transfected with pcDNA3.1(+)/RIZ1 and pcDNA3.1(+).Changes in gene expression profile were screened and the microarray results were confirmed by reverse transcriptionpolymerase chain reaction(RT-PCR).Nude mice were inoculated with TE13 cells to establish ESCC xenografts.After two weeks,the inoculated mice were randomly divided into three groups.Tumors were injected with normal saline,transfection reagent pcDNA3.1(+)and transfection reagent pcDNA3.1(+)/RIZ1,respectively.Tumor development was quantified,and changes in gene expression of RIZ1 transfected tumors were detected by RT-PCR and Western blotting.RESULTS:DNA microarray data showed that RIZ1transfection induced widespread changes in gene expression profile of cell line TE13,with 960 genes upregulated and 1163 downregulated.Treatment of tumor xenografts with RIZ1 recombinant plasmid significantly inhibited tumor growth,decreased tumor size,and increased expression of RIZ1 mRNA compared to control groups.The changes in gene expression profile were also observed in vivo after RIZ1 transfection.Most of the differentially expressed genes were associated with cell development,supervision of viral replication,lymphocyte costimulatory and immune system development in esophageal cells.RIZ1 gene may be involved in multiple cancer pathways,such as cytokine receptor interaction and transforming growth factor beta signaling.CONCLUSION:The development and progression of esophageal cancer are related to the inactivation of RIZ1.Virus infection may also be an important factor.展开更多
目的:探讨小脑锌指结构1(zinc finger protein of the cerebellum 1,ZIC1)基因在人子宫内膜癌组织中的表达情况及其与临床参数之间的关系和预后预测价值。方法:收集江苏大学附属昆山医院及上海市第六人民医院自2008年1月至2011年12月收...目的:探讨小脑锌指结构1(zinc finger protein of the cerebellum 1,ZIC1)基因在人子宫内膜癌组织中的表达情况及其与临床参数之间的关系和预后预测价值。方法:收集江苏大学附属昆山医院及上海市第六人民医院自2008年1月至2011年12月收治的子宫内膜癌患者43例,取肿瘤组织及相应的距原发灶边缘2 cm以上的癌旁黏膜组织标本。应用免疫组织化学法、Western blotting及RT-PCR法检测43例子宫内膜癌原发灶组织及癌旁黏膜组织中ZIC1的表达,分析其与临床病理参数之间的关系,并通过5年生存情况分析评价其预后价值。结果:子宫内膜癌组织中ZIC1的m RNA水平和蛋白水平均明显高于癌旁组织。ZIC1蛋白在子宫内膜癌组织中呈现高表达,其阳性表达率为72.1%(31/43),在癌旁组织中阳性表达率为39.5%(17/43)。ZIC1 m RNA的表达与淋巴结转移及FIGO分期有关(P<0.01或P<0.05)。ZIC1低表达患者的5年生存率明显高于高表达患者(66.7%vs 38.7%,P<0.05)。单因素分析发现ZIC1(高表达vs低表达)风险比(HR)为2.66,95%CI:1.07-5.89,P=0.047;经多因素分析调整后,HR=2.25,95%CI:1.36-3.71,P=0.002,说明高水平的ZIC1与子宫内膜癌术后预后不良密切相关。结论:ZIC1表达水平的升高在子宫内膜癌的发生发展中起着重要作用,可能成为子宫内膜癌治疗和预后测评的一个新的指标。展开更多
目的:探讨小脑锌指结构1(zinc finger of the cerebellum 1,ZIC1)基因在人乳腺癌组织中的表达及其对乳腺癌MDA-MB-231细胞增殖能力的影响。方法:应用免疫组织化学法检测238例乳腺癌和40例癌旁组织中ZIC1蛋白的表达,并分析其与临床病理...目的:探讨小脑锌指结构1(zinc finger of the cerebellum 1,ZIC1)基因在人乳腺癌组织中的表达及其对乳腺癌MDA-MB-231细胞增殖能力的影响。方法:应用免疫组织化学法检测238例乳腺癌和40例癌旁组织中ZIC1蛋白的表达,并分析其与临床病理参数之间的关系。构建真核表达载体pc DNA3.1-ZIC1,通过脂质体法瞬时转染乳腺癌MDA-MB-231细胞(实验组),以未转染细胞和转染pc DNA3.1空质粒细胞分别作为空白对照和阴性对照。采用反转录PCR及蛋白质印迹法检测3组细胞中ZIC1 mRNA及蛋白的表达,采用MTT法检测细胞增殖能力的变化。结果:ZIC1蛋白在乳腺癌组织中的阳性表达率(39.1%,93/238)明显低于癌旁组织(77.5%,31/40),两者差异有统计学意义(P<0.05)。乳腺癌组织中ZIC1蛋白的表达与病理分型、组织学分级和淋巴结转移相关(P<0.05)。ZIC1 mRNA和蛋白在实验组中均有表达,在两对照组中不表达;与阴性对照组和空白对照组相比,实验组细胞的增殖能力明显减弱。结论:乳腺癌组织中ZIC1蛋白的阳性表达率明显下降并与多项临床病理参数相关,转染ZIC1基因后能够明显抑制乳腺癌细胞的增殖,提示ZIC1基因在乳腺癌的发生、发展中发挥重要作用。展开更多
文摘The myeloid zinc finger gene-1 (MZF-1) encodes a putative transcription factor whose expression has been implicated in myeloid differentiation. To study the role of the nMZF-1 in myploid differentiation,we characterized MZF-1 protein expr.ession,cellular localization,and phosphorylation in leukemia cell lines and leukemia cells.MZF1 protein expression was found only in myeloid cells. In proliferating HL-60 cells,MZF-1 was localized to the nucleus with some cytoplasmic distribution; however,upon retinoic acid (RA)induced granulocytic differentiation, MZF-1 became restricted to the nucleus.In32 PO4-la labelled HL-60 cell, MZF-1 was immunoprecipitated as a phosphoprotein doublet of 53￣54kDa. MZF-1 phosphorylation increased after acute stimulation of HL-60 with granulocytemacrophage colony stimulating factor (GM-CSF), interleukin-3(IL-3),phorbol ester,and serum.Chronic GM-CSf treatment of HL-60 cells potentiating granulocytic differentiation sustained the hyperphosphorylated state of MZF-1,whereas chronic treatment with TPA leading to monocytic-macrophage differentiation was accompanied by the disappearance of the 53 kDa MZF-1 phosphoprotein and the appearance of cross-reactive 69 and 105kDa phosphoprotein species. K562 human myeloblastic cells which are resistant to granulocytic differentiation express both the 53 kDa MZF-1 protein and the cross reactive 69 and 105 kDa proteins,but the 53 kDa MZF-1 protein is not detectable phosphorylated under any experimental conditions. Acute promyelocytic leukemic cells exhibited the 53kDa phosphoprotein,whereas monocytic leukemia cells expressed only the 69 and 105 kDa MZF-1 related phosphoproteins. The studies demonstrate that MZF-1 is a nuclear protein whose phosphorylation is associated with the granulocytic commitment of myeloid cells.
基金Supported by Grant-in-aid from Specialized Research Fund for the Doctoral Program of Higher Education,No. 20091202110009Grant-in-aid from Natural Science Foundation of Tianjin,No. 10JCYBJC11300
文摘AIM： To investigate the promoter region methylation status of retinoblastoma protein-interacting zinc finger gene 1 （RIZ1） in the human esophageal squamous cell carcinoma （ESCC） cell lines and tissues and verify the relationship between methylation of RIZ1 and oncogen- esis, tumor progression and metastasis etc of ESCC. METHODS： Methylation-specific polymerase chain reac- tion （MSP） was used to investigate the promoter region methylation status of RIZ1 in 6 ESCC cell lines. One cell line where RIZ1 promoter region methylation was de- tected was selected for the next study, where the cell line was treated with 5-aza-CdR. Real-time polymerase chain reaction was used to investigate its influence on the transcription of RIZ1. Experiments using frozenpathological specimens from 47 ESCC patients were performed using the same MSP methodology. RESULTS： Promoter methylation of RIZ1 gene was detected in TEl3, CaEs17 and EC109 cell lines and the cell line TEl3 was chosen for further study. The expression of RIZl mRNA in TE-13 was up-regulated after treatment with 5-aza-CdR. The rate of methyla- tion in carcinomas tissues was significantly higher than those in matched neighboring normal and distal ending normal tissue, and the deviation of data was statisti- cally significant （2,2 = 24.136, P 〈 0.01）. Analysis of the gender, age familial history, tumour deviation, tumour saturation, lymph gland displacement and clinical stag- ing of 47 samples from ESCC patients showed that the fluctuation of data was not statistically significant. CONCLUSION： Promoter methylation may play an im- portant role in the epigenetic silencing of RIZ1 gene expression in human ESCC. RIZ1 is considered to be a potential tumor suppressor gene and may be a biologi- cal parameter for testing early stage human ESCC.
基金Supported by The National Natural Science Foundation of China,No.81201945Science Foundation of Tianjin Medical University,No.2011KY08+1 种基金Doctoral Program of Higher Education Research Fund,No.20091202110009Natural Science Foundation of Tianjin,China,No.10JCYBJC11300
文摘AIM:To investigate the effect of retinoblastoma protein-interacting zinc finger gene 1(RIZ1)upregulation in gene expression profile and oncogenicity of human esophageal squamous cell carcinoma(ESCC)cell line TE13.METHODS:TE13 cells were transfected with pcDNA3.1(+)/RIZ1 and pcDNA3.1(+).Changes in gene expression profile were screened and the microarray results were confirmed by reverse transcriptionpolymerase chain reaction(RT-PCR).Nude mice were inoculated with TE13 cells to establish ESCC xenografts.After two weeks,the inoculated mice were randomly divided into three groups.Tumors were injected with normal saline,transfection reagent pcDNA3.1(+)and transfection reagent pcDNA3.1(+)/RIZ1,respectively.Tumor development was quantified,and changes in gene expression of RIZ1 transfected tumors were detected by RT-PCR and Western blotting.RESULTS:DNA microarray data showed that RIZ1transfection induced widespread changes in gene expression profile of cell line TE13,with 960 genes upregulated and 1163 downregulated.Treatment of tumor xenografts with RIZ1 recombinant plasmid significantly inhibited tumor growth,decreased tumor size,and increased expression of RIZ1 mRNA compared to control groups.The changes in gene expression profile were also observed in vivo after RIZ1 transfection.Most of the differentially expressed genes were associated with cell development,supervision of viral replication,lymphocyte costimulatory and immune system development in esophageal cells.RIZ1 gene may be involved in multiple cancer pathways,such as cytokine receptor interaction and transforming growth factor beta signaling.CONCLUSION:The development and progression of esophageal cancer are related to the inactivation of RIZ1.Virus infection may also be an important factor.
文摘目的:探讨小脑锌指结构1(zinc finger protein of the cerebellum 1,ZIC1)基因在人子宫内膜癌组织中的表达情况及其与临床参数之间的关系和预后预测价值。方法:收集江苏大学附属昆山医院及上海市第六人民医院自2008年1月至2011年12月收治的子宫内膜癌患者43例,取肿瘤组织及相应的距原发灶边缘2 cm以上的癌旁黏膜组织标本。应用免疫组织化学法、Western blotting及RT-PCR法检测43例子宫内膜癌原发灶组织及癌旁黏膜组织中ZIC1的表达,分析其与临床病理参数之间的关系,并通过5年生存情况分析评价其预后价值。结果:子宫内膜癌组织中ZIC1的m RNA水平和蛋白水平均明显高于癌旁组织。ZIC1蛋白在子宫内膜癌组织中呈现高表达,其阳性表达率为72.1%(31/43),在癌旁组织中阳性表达率为39.5%(17/43)。ZIC1 m RNA的表达与淋巴结转移及FIGO分期有关(P<0.01或P<0.05)。ZIC1低表达患者的5年生存率明显高于高表达患者(66.7%vs 38.7%,P<0.05)。单因素分析发现ZIC1(高表达vs低表达)风险比(HR)为2.66,95%CI:1.07-5.89,P=0.047;经多因素分析调整后,HR=2.25,95%CI:1.36-3.71,P=0.002,说明高水平的ZIC1与子宫内膜癌术后预后不良密切相关。结论:ZIC1表达水平的升高在子宫内膜癌的发生发展中起着重要作用,可能成为子宫内膜癌治疗和预后测评的一个新的指标。
文摘目的:探讨小脑锌指结构1(zinc finger of the cerebellum 1,ZIC1)基因在人乳腺癌组织中的表达及其对乳腺癌MDA-MB-231细胞增殖能力的影响。方法:应用免疫组织化学法检测238例乳腺癌和40例癌旁组织中ZIC1蛋白的表达,并分析其与临床病理参数之间的关系。构建真核表达载体pc DNA3.1-ZIC1,通过脂质体法瞬时转染乳腺癌MDA-MB-231细胞(实验组),以未转染细胞和转染pc DNA3.1空质粒细胞分别作为空白对照和阴性对照。采用反转录PCR及蛋白质印迹法检测3组细胞中ZIC1 mRNA及蛋白的表达,采用MTT法检测细胞增殖能力的变化。结果:ZIC1蛋白在乳腺癌组织中的阳性表达率(39.1%,93/238)明显低于癌旁组织(77.5%,31/40),两者差异有统计学意义(P<0.05)。乳腺癌组织中ZIC1蛋白的表达与病理分型、组织学分级和淋巴结转移相关(P<0.05)。ZIC1 mRNA和蛋白在实验组中均有表达,在两对照组中不表达;与阴性对照组和空白对照组相比,实验组细胞的增殖能力明显减弱。结论:乳腺癌组织中ZIC1蛋白的阳性表达率明显下降并与多项临床病理参数相关,转染ZIC1基因后能够明显抑制乳腺癌细胞的增殖,提示ZIC1基因在乳腺癌的发生、发展中发挥重要作用。
