Abstract
Purpose
To identify methylation-silenced genes in acute myeloid leukemia (AML).
Methods
Microarray analyses were performed in AML cell line HL-60 cells exposed to the demethylating agent 5-aza-2dC. The methylation status and expression of glioma pathogenesis-related protein 1 (GLIPR1), one of highly induced genes by demethylation, were further detected in six hematopoietic malignancy cell lines and 260 bone marrow samples from leukemia patients and nonmalignant diseases as control, as well as pre-treated and post-treated bone marrow samples from 24 complete remission AML patients received chemotherapy using MS-PCR, bisulfite DNA sequencing, RT-PCR, and Western blotting.
Results
One hundred and nine genes were significantly induced by demethylation in HL-60 cells, 12 genes of which were confirmed by RT-PCR. GLIPR1, a tumor suppressor gene, was frequently methylation-silenced in AML cell lines and AML patients, but not in the other hematopoietic malignancy cell lines and patients. The frequencies of methylation-silenced GLIPR1 in the pre-treatment were significantly higher than those in the post-treatment in complete remission AML patients.
Conclusion
We identify 109 genes induced by demethylation in HL-60 cells, and demonstrate that GLIPR1 is a methylation-silenced gene in the AML patients, and may serve as a marker for monitoring disease activity during therapy in the AML patients. The data provide the important information for studying the pathogenesis of AML and discovering the target genes of methylating agents.
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Acknowledgments
This work was supported by National Nature Science Foundation of China (30973290), Outstanding Scholars of New Era from Ministry of Education of China (2002-48), Lotus Scholars Program of Hunan Province, China (2007-362), Key research program from Science and Technology Committee of Hunan Province, China (2010FJ2009), and Aid program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province, China.
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No conflicts of interest were declared.
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Yan-Hua Xiao, Xin-Hui Li and Tan Tan contributed equally to this work.
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Xiao, YH., Li, XH., Tan, T. et al. Identification of GLIPR1 tumor suppressor as methylation-silenced gene in acute myeloid leukemia by microarray analysis. J Cancer Res Clin Oncol 137, 1831–1840 (2011). https://doi.org/10.1007/s00432-011-1065-2
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DOI: https://doi.org/10.1007/s00432-011-1065-2