Summary
The molecular pathogenesis of leukemia is poorly understood. Earlier studies have shown both Wilms’ tumor 1 suppressor gene (WT1) and CML28 abnormally expressed in malignant diseases of the hematopoietic system and WT1 played an important role in leukemogenesis. However, the relationship between molecular CML28 and WT1 has not been reported. Here we described the use of small interfering RNA (siRNA) against WT1 and CML28 in leukemic cell line K562 to examine the interaction between CML28 and WT1. WT1 and CML28 gene expression in transfected K562 cells was detected by using RQ-PCR and Western blotting. K562 cells transfected with WT1-siRNA could greatly decrease both mRNA and protein expression levels of WT1 and CML28. In contrast, CML28-siRNA did not exert effect on WT1. Further, subcellular co-localization assay showed that the two proteins could co-localize in the cytoplasm of K562 cells, but WT1/CML28 complexes were not detected by using immunoprecipitation. It was suggested that there exists the relationship between CML28 and WT1. CML28 may be a downstream target molecule of WT1 and regulated by WT1, which will provide important clues for further study on the role of CML28 and WT1 in leukemic cells.
Similar content being viewed by others
References
Buckler AJ, Pelletier J, Haber DA, et al. Isolation, characterization, and expression of the murine Wilms’ tumor gene (WT1) during kidney development. Mol Cell Biol, 1991,11(3):1707–1712
Rauscher FJ 3rd, Morris JF, Tournay OE, et al. Binding of the Wilms’ tumor locus zinc finger protein to the EGR-1 consensus sequence. Science, 1990,250(4985): 1259–1262
Dey BR, Sukhatme VP, Roberts AB, et al. Repression of the transforming growth factor-beta 1 gene by the Wilms’ tumor suppressor WT1 gene product. Mol Endocrinol, 1994,8(5):595–602
Loeb DM. WT1 influences apoptosis through transcriptional regulation of Bcl-2 family members. Cell Cycle, 2006,5(12):1249–1253
Oh S, Song Y, Yim J, et al. The Wilms’ tumor 1 tumor suppressor gene represses transcription of the human telomerase reverse transcriptase gene. J Biol Chem, 1999,274(52):37 473–37 478
Cilloni D, Gottardi E, De Micheli D, et al. Quantitative assessment of WT1 expression by real time quantitative PCR may be a useful tool for monitoring minimal residual disease in acute leukemia patients. Leukemia, 2002,16(10):2115–2121
Inoue K, Sugiyama H, Ogawa H, et al. WT1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood, 1994,84(9):3071–3079
Brieger J, Weidmann E, Maurer U, et al. The Wilms’ tumor gene is frequently expressed in acute myeloblastic leukemias and may provide a marker for residual blast cells detectable by PCR. Ann Oncol, 1995,6(8):811–816
Bergmann L, Miething C, Maurer U, et al. High levels of Wilms’ tumor gene (wt1) mRNA in acute myeloid leukemias are associated with a worse long-term outcome. Blood, 1997,90(3):1217–1225
Barragan E, Cervera J, Bolufer P, et al. Prognostic implications of Wilms’ tumor gene (WT1) expression in patients with de novo acute myeloid leukemia. Haematologica, 2004,89(8):926–933
Chiusa L, Francia di Celle P, Campisi P, et al. Prognostic value of quantitative analysis of WT1 gene transcripts in adult acute lymphoblastic leukemia. Haematologica, 2006,91(2):270–271
Huff V, Saunders GF. Wilms tumor genes. Biochim Biophys Acta, 1993,1155(3):295–306
Call KM, Glaser T, Ito CY, et al. Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms’ tumor locus. Cell, 1990,60(3): 509–520
Yamagami T, Sugiyama H, Inoue K, et al. Growth inhibition of human leukemic cells by WT1 (Wilms’ tumor gene) antisense oligodeoxynucleotides: implications for the involvement of WT1 in leukemogenesis. Blood, 1996,87(7):2878–2884
Yang XF, Wu CJ, Chen L, et al. CML28 is a broadly immunogenic antigen, which is overexpressed in tumor cells. Cancer Res, 2002,62(19):5517–5522
Guo X, Ma J, Sun J, et al. The zinc-finger antiviral protein recruits the RNA processing exosome to degrade the target mRNA. Proc Natl Acad Sci USA, 2007,104(1): 151–156
Yang XF, Wu CJ, McLaughlin S, et al. CML66, a broadly immunogenic tumor antigen, elicits a humoral immune response associated with remission of chronic myelogenous leukemia. Proc Natl Acad Sci USA, 2001,98(13): 7492–7497
Wu CJ, Biernacki M, Kutok JL, et al. Graft-versus-leukemia target antigens in chronic myelogenous leukemia are expressed on myeloid progenitor cells. Clin Cancer Res, 2005,11(12):4504–4511
Zhou H, Zhang D, Wang Y, et al. Induction of CML28-specific cytotoxic T cell responses using co-transfected dendritic cells with CML28 DNA vaccine and SOCS1 small interfering RNA expression vector. Biochem Biophys Res Commun, 2006,347(1):200–207
Zhang DH, Zhou HS, Wang YY, et al. Construction and expression of dendritic cell nucleic acid vaccine containing CML28 gene in human dendritic cells. Zhongguo Shi Yan Xue Ye Xue Za Zhi (Chinese), 2005,13(4):631–636
Zhang DH, Dai M, Zhou HS, et al. Monitoring CML28 mRNA levels in patients before and after HSCT by real-time quantitative RT-PCR. Zhongguo Shi Yan Xue Ye Xue Za Zhi (Chinese), 2005,13(5):843–847
Bai XL, Mao X, Zhang B, et al. The role of CML28 in regulation of proliferation and apoptosis of K562 cell. Med J Chin PAPF, 2012,23(8):678–681
Jacobsohn DA, Tse WT, Chaleff S, et al. High WT1 gene expression before haematopoietic stem cell transplant in children with acute myeloid leukaemia predicts poor event-free survival. Br J Haematol, 2009,146(6):669–674
Glienke W, Maute L, Koehl U, et al. Effective treatment of leukemic cell lines with wt1 siRNA. Leukemia, 2007,21(10):2164–2170
Gao F, Maiti S, Alam N, et al. The Wilms tumor gene, Wt1, is required for Sox9 expression and maintenance of tubular architecture in the developing testis. Proc Natl Acad Sci USA, 2006,103(32):11 987–11 992
Martínez-Estrada OM, Lettice LA, Essafi A, et al. Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin. Nature Genet, 2010,42(1):89–93
Chen J, Santillan DA, Koonce M, et al. Loss of MLL PHD finger 3 is necessary for MLL-ENL-induced hematopoietic stem cell immortalization. Cancer Res, 2008,68(15): 6199–6207
Rokudai S, Aikawa Y, Tagata Y, et al. Monocytic leukemia zinc finger (MOZ) interacts with p53 to induce p21 expression and cell-cycle arrest. J Biol Chem, 2009,284(1):237–244
Morrison AA, Viney RL, Ladomery MR. The post-trans-criptional roles of WT1, a multifunctional zinc-finger protein. Biochim Biophys Acta, 2008,1785(1): 55–62
Bevilacqua A, Ceriani MC, Capaccioli S, et al. Post-transcriptional regulation of gene expression by degradation of messenger RNAs. J Cell Physiol, 2003, 195(3):356–372
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mao, X., Zhang, B., Liu, Ll. et al. Interaction of Human Genes WT1 and CML28 in Leukemic Cells. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 33, 37–42 (2013). https://doi.org/10.1007/s11596-013-1068-0
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11596-013-1068-0