锌指蛋白ZNF185对粒细胞白血病细胞增殖的影响及表达调控研究
|
摘要
目的检测锌指蛋白ZNF185对粒细胞白血病细胞增殖的影响,探讨调控ZNF185表达的分子机制。方法采用Western-blot检测ZNF185的表达。克隆ZNF185编码序列,转染粒细胞白血病细胞,检测细胞增殖。采用甲基化特异PCR检测ZNF185基因的甲基化。结果 ZNF185在人正常中性粒细胞中高表达,而在粒细胞白血病细胞中表达显著降低;过表达ZNF185显著抑制白血病细胞增殖;ZNF185在粒细胞白血病细胞中的甲基化显著增加。结论 DNA甲基化抑制ZNF185在白血病细胞中的表达进而促进细胞增殖。
Objective To observe the effects of Zinc-finger protein 185(ZNF185)on cell proliferation of granulocytic leukemia,and to explore the molecular mechanism of regulating expression of ZNF185.Methods The expression of ZNF185 was detected by Western Blot.Coding sequence of ZNF1 85 gene was cloned and transfected into granulocytic leukemia cell lines,and the cell proliferation was detected by means of cell counting.Methylation specific PCR(MSP)was used to detect methylation condition of ZNF185.Results ZNF185 protein was highly expressed in normal neutrophils,however,the expression of ZNF185 was obviously decreased in granulocytic leukemia cell line.The overexpression of ZNF185 significantly inhibited the proliferation of leukemia cells.The methylation of ZNF185 was remarkably increased in granulocytic leukemia cells.Conclusion DNA methylation inhibits the expression of ZNF185 in leukemia cells,as a result,which promotes the cell proliferation.
Objective To observe the effects of Zinc-finger protein 185(ZNF185)on cell proliferation of granulocytic leukemia,and to explore the molecular mechanism of regulating expression of ZNF185.Methods The expression of ZNF185 was detected by Western Blot.Coding sequence of ZNF1 85 gene was cloned and transfected into granulocytic leukemia cell lines,and the cell proliferation was detected by means of cell counting.Methylation specific PCR(MSP)was used to detect methylation condition of ZNF185.Results ZNF185 protein was highly expressed in normal neutrophils,however,the expression of ZNF185 was obviously decreased in granulocytic leukemia cell line.The overexpression of ZNF185 significantly inhibited the proliferation of leukemia cells.The methylation of ZNF185 was remarkably increased in granulocytic leukemia cells.Conclusion DNA methylation inhibits the expression of ZNF185 in leukemia cells,as a result,which promotes the cell proliferation.
引文
1Heiss NS,Gloeckner G,Bchner D,et al.Genomic structure of a novel LIM domain gene(ZNF185)in Xq28and comparisons with the ortholo-gous murine transcript.Genomics,1997,43:329-338.
2Rabbitts TH,Boehm T.LIM domains.Nature,1990,346:418.
3Sanchez-Garcia I,Rabbitts T.The LIM domains:a new structural motif found in zinc-finger-like protein.Trends Genet,1994,10:315-320.
4Zhang JS,Gong A,Young CY.ZNF185,an actin-cytoskeletonassociated growth inhibitory LIM protein in prostate cancer.Oncogene,2007,26:111-122.
5Gonzalez H,Gujrati M,Frederick M,et al.Identification of9genes differentially expressed in head and neck squamous cell carcinoma.Arch Oto-laryngol Head Neck Surg,2003,129:754-759.
6Vanaja DK,Cheville JC,Iturria SJ,et al.Transcriptional silencing of zinc finger protein185identified by expression profiling is associated with prostate cancer progression.Cancer Res,2003,63:3877-3882.
7Medina PP,Carretero J,Ballestar E,et al.Sanchez-Cespedes M.Transcriptional targets of the chromatin-remodelling factor SMARCA4/BRG1in lung cancer cells.Hum Mol Genet,2005,14:973-982.
8Chomel JC,Sorel N,Guilhot J,et al.BCR-ABL expression in leukemic progenitors and primitive stem cells of patients with chronic myeloid leu-kemia.Blood,2012,119:2964-2965.
9Comb M,Goodman HM.CpG methylation inhibits proenkephalin gene expression and binding of the transcription factor AP-2.Nucleic Acids Res,1990,18:3975-3982.
2Rabbitts TH,Boehm T.LIM domains.Nature,1990,346:418.
3Sanchez-Garcia I,Rabbitts T.The LIM domains:a new structural motif found in zinc-finger-like protein.Trends Genet,1994,10:315-320.
4Zhang JS,Gong A,Young CY.ZNF185,an actin-cytoskeletonassociated growth inhibitory LIM protein in prostate cancer.Oncogene,2007,26:111-122.
5Gonzalez H,Gujrati M,Frederick M,et al.Identification of9genes differentially expressed in head and neck squamous cell carcinoma.Arch Oto-laryngol Head Neck Surg,2003,129:754-759.
6Vanaja DK,Cheville JC,Iturria SJ,et al.Transcriptional silencing of zinc finger protein185identified by expression profiling is associated with prostate cancer progression.Cancer Res,2003,63:3877-3882.
7Medina PP,Carretero J,Ballestar E,et al.Sanchez-Cespedes M.Transcriptional targets of the chromatin-remodelling factor SMARCA4/BRG1in lung cancer cells.Hum Mol Genet,2005,14:973-982.
8Chomel JC,Sorel N,Guilhot J,et al.BCR-ABL expression in leukemic progenitors and primitive stem cells of patients with chronic myeloid leu-kemia.Blood,2012,119:2964-2965.
9Comb M,Goodman HM.CpG methylation inhibits proenkephalin gene expression and binding of the transcription factor AP-2.Nucleic Acids Res,1990,18:3975-3982.