Molecular features and expression of DAZAP2 in human multiple myeloma  

作　　者：SHI Yi-wu SHEN Rong REN Wei TANG Li-jun TAN Da-ren HU Wei-xin 

机构地区：[1]Molecular Biology Research Center, School of Biological Science and Technology, Central South University, Changsha 410078, China [2]Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China

出　　处：《Chinese Medical Journal》2007年第19期1659-1665,共7页中华医学杂志（英文版）

基　　金：This work was supported by the grants from the National Natural Science Foundation of China(No.30270750, 30300406 and 30400529).

摘　　要：Background In our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA （cDNA） chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma （MM） patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM. Methods The full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples. Results DAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C （PKC） phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients （P〈0.001） that matched with the results of the RT-PCR. Conclusions DAZAP2 is downregulated in MM samples and itBackground In our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA （cDNA） chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma （MM） patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM. Methods The full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples. Results DAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C （PKC） phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients （P〈0.001） that matched with the results of the RT-PCR. Conclusions DAZAP2 is downregulated in MM samples and it

关 键 词：DAZAP2 gene structure gene expression multiple myeloma 

分 类 号：R738.1[医药卫生—肿瘤]