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[慢性髓性白血病急变期相关蛋白的初步研究]

[Preliminary study of proteins related to blast crisis in chronic myeloid leukemia].

作者信息

Zhu Hong-qian, Liu Xiao-li, Li Rong, Du Qing-feng, Zhang Song, Yao Feng, Liu Zhi

机构信息

Department of Hematology, Nanfang Hospital, Nanfang Medical College, Guangzhou, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2009 Sep;31(9):655-9.

Abstract

OBJECTIVE

To identify and compare the expression profiles of differential proteins between chronic phase and blast crisis in chronic myeloid leukemia (CML) by proteomic analysis, and screen the proteins related to blast crisis.

METHODS

The total cellular proteins from the bone marrow cells at chronic phase (CP) and blast crisis (BC) in CML were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and analyzed with ImageMaster 5.0 software to screen the differential protein spots. Differential protein spots were identified by mass spectrometry for peptide mass fingerprint in combination with database searching from SWISS-PROT. Then 3 protein spots were selected to verify at protein and mRNA levels by Western blot and semi-quantitative RT-PCR, separately.

RESULTS

Comparing gel pages from CML-CP and CML-BC, the expression of 13 protein spots decreased and 25 protein spots increased significantly in CML-BC. Twenty differential protein spots were identified by mass spectrometry and 15 were successfully determined. The results of Western blotting were similar to those of 2-DE and showed a high expression of hnRNPK, annexin A1 and RhoA. Semi-quantitative RT-PCR analysis showed that there was no correlation between the protein expression changes and mRNA levels of hnRNPK, annexin A1 and RhoA.

CONCLUSION

A group of proteins associated with blast crisis are obtained and the results may provide clues for further research to elucidate the role of these proteins in CML-BC carcinogenesis and to develop potential associated biomarkers.

摘要

目的

通过蛋白质组学分析鉴定并比较慢性髓性白血病(CML)慢性期与急变期差异蛋白的表达谱,筛选与急变期相关的蛋白。

方法

采用二维聚丙烯酰胺凝胶电泳(2-DE)分离CML慢性期(CP)和急变期(BC)骨髓细胞的总细胞蛋白,并用ImageMaster 5.0软件进行分析,以筛选差异蛋白点。通过质谱结合SWISS-PROT数据库搜索对差异蛋白点进行肽质量指纹图谱鉴定。然后分别选择3个蛋白点通过蛋白质免疫印迹法(Western blot)和半定量逆转录-聚合酶链反应(RT-PCR)在蛋白和mRNA水平进行验证。

结果

比较CML-CP和CML-BC的凝胶图谱,CML-BC中有13个蛋白点表达下降,25个蛋白点表达显著增加。通过质谱鉴定出20个差异蛋白点,成功鉴定出15个。蛋白质免疫印迹结果与2-DE结果相似,显示hnRNPK、膜联蛋白A1和RhoA高表达。半定量RT-PCR分析表明,hnRNPK、膜联蛋白A1和RhoA的蛋白表达变化与mRNA水平之间无相关性。

结论

获得了一组与急变期相关的蛋白,这些结果可能为进一步研究阐明这些蛋白在CML-BC致癌过程中的作用以及开发潜在相关生物标志物提供线索。

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