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Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Dec;21(6):1394-8. doi: 10.7534/j.issn.1009-2137.2013.06.006.

[Effect of AML1-ETO fusion protein on the expression of BCL-2].

[Article in Chinese]

Author information

  • 1Medical Ecsomatics College of Beihua University, Jilin 132013, Jilin Province, China.
  • 2Department of Laboratorial Examination, Jilin Medical College, Jilin 132013, Jilin Province, China.
  • 3Cyrus Tang Hematology Center of Soochow University, Suzhou 215000, Jiangsu Province, China.
  • 4Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou 215000, Jiangsu Province, China.
  • 5Department of Cell Biology, School of Biology & Basic Medical Science, Soochow University, Suzhou 215000, Jiangsu Province, China.
  • 6Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou 215000, Jiangsu Province, China. E-mail: szchenzx@ 263.net.

Abstract

This study was aimed to investigate the effect of AML1-ETO fusion protein on the anti-apoptotic gene BCL-2 in leukemic cells and to explore its role in leukemogenesis. The apoptotic levels of U937-WT, U937-Mock and U937-A/E1-4 cells were examined by flow cytometry. And cleaved caspase-3 protein expression was detected by Western blot. BCL-2 gene expression both in AML1-ETO-expressing cells or U937 nonexpressing cells and in leukemia cells of AML patients with or without t(8;21) was assessed by quantitative PCR. The chromatin immunoprecipitation (ChIP)-based PCR was used to investigate the direct interaction between the AML1-ETO and BCL-2 promoter in AML1-ETO positive leukemia cell line. The results indicated that in U937-A/E cells but not in U937-WT or U937-Mock cells, apoptotic cells statistically significantly increased, and AML1-ETO expression also significantly enhanced activation of caspase-3. AML1-ETO-expressing cell subclones displayed significantly low levels of BCL-2 mRNA in comparison with the non-transfected U937. In primary bone marrow cells of acute myeloid leukemia containing AML1-ETO, levels of BCL-2 mRNA were markedly lower as compared with other acute myeloid leukemias lacking this translocation. The enriched regions in transfected cells were located within BCL-2 promoter. It is concluded that BCL-2 is the direct target gene of AML1-ETO. AML1-ETO can down-regulate the expression of BCL-2.

PMID:
24370018
[PubMed - indexed for MEDLINE]
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