Identification of GLIPR1 tumor suppressor as methylation-silenced gene in acute myeloid leukemia by microarray analysis

• Published in: Journal of cancer research and clinical oncology 2011-12, Vol.137 (12), p.1831-1840
• Main Authors: Xiao, Yan-Hua, Li, Xin-Hui, Tan, Tan, Liang, Ting, Yi, Hong, Li, Mao-Yu, Zeng, Gu-Qing, Wan, Xun-Xun, Qu, Jia-Quan, He, Qiu-Yan, Li, Jian-Huang, Chen, Yu, Xiao, Zhi-Qiang
• Format: Article
• Language: English
• Subjects: Acute myeloid leukemia; Adolescent; Adult; Aged; Antineoplastic agents; Biological and medical sciences; Bisulfite; Bone marrow; Brain tumors; Cancer Research; Cell Line, Tumor; Chemotherapy; Data processing; Demethylation; DNA Methylation; DNA sequencing; Female; Gene Silencing; Genes; Genes, Tumor Suppressor; Glioma; Hematologic and hematopoietic diseases; Hematology; Hemopoiesis; Humans; Internal Medicine; Leukemia; Leukemia, Myeloid, Acute - genetics; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Male; Malignancy; Medical sciences; Medicine; Medicine & Public Health; Middle Aged; Neoplasm Proteins - genetics; Nerve Tissue Proteins - genetics; Oligonucleotide Array Sequence Analysis - methods; Oncology; Original Paper; Pathogenesis; Pathogenesis-related proteins; Pharmacology. Drug treatments; Polymerase chain reaction; Promoter Regions, Genetic; Remission; Reverse Transcriptase Polymerase Chain Reaction; Tumor suppressor genes; Western blotting
• ISSN: 0171-5216; 1432-1335
• DOI: 10.1007/s00432-011-1065-2

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.