Breast cancer, which is the most common type of cancer among women, is a heterogenous disease. It results from progressive accumulation of genetic and epigenetic alterations in different genes. The Dok1 protein has been identified as the major substrate of protein tyrosine kinases in hematopoietic cells. It is considered as a tumor suppressor due to the reports which describe its inhibitory effect on major oncogenic signaling pathways such as Mek/Erk/PI3k/Akt and Wnt/β-catenin. In this study, we investigated the mutation frequency of the DOK1 gene in 118 breast tumors using Sanger sequencing and DOK1 mRNA expression level in 63 breast cancer samples using qRT-PCR methods. Although the mutation frequency was low DOK1 mRNA expression levels were significantly reduced (63.5%) in the tumors compared to adjacent non-cancerous tissue. We also correlated expression changes with clinicopathological characteristics. Low mRNA levels correlated with age (p = 0.01) and c-erbB-2 (p = 0.05). In most of the previous reports, down-regulation of DOK1 mRNA expression has been associated with promoter methylation. We identified four different coding sequence alterations in 5.1% (6/118) of the tumor samples. However, all of these alterations were located in the functional domains of the protein. Therefore, these mutations may affect the function and/or cellular localization of the protein and contribute to cancer progression by this way. In conclusion our data indicate that DOK1 acts as a tumor suppressor in breast cancer and association of Dok1 with the c-erbB-2 mediated mechanism of action in breast cancer needs to be investigated.
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Yamanashi Y, Baltimore D (1997) Identification of the Abl- and rasGAP-associated 62 kDa protein as a docking protein, Dok. Cell 88:205–211
Carpino N, Wisniewski D, Strife A et al (1997) p62(dok): a constitutively tyrosine-phosphorylated, GAP-associated protein in chronic myelogenous leukemia progenitor cells. Cell 88:197–204
Nelms K, Snow AJ, Noben-Trauth K (1998) Dok1 encoding p62(dok) maps to mouse chromosome 6 and human chromosome 2 in a region of translocation in chronic lymphocytic leukemia. Genomics 53:243–245
Mashima R, Hishida Y, Tezuka T et al (2009) The roles of Dok family adapters in immunoreceptor signaling. Immunol Rev 232:273–285
Niki M, Di Cristofano A, Zhao M et al (2004) Role of Dok-1 and Dok-2 in leukemia suppression. J Exp Med 200:1689–1695
Yasuda T, Shirakata M, Iwama A et al (2004) Role of Dok-1 and Dok-2 in myeloid homeostasis and suppression of leukemia. J Exp Med 200:1681–1687
Cong F, Yuan B, Goff SP (1999) Characterization of a novel member of the DOK family that binds and modulates Abl signaling. Mol Cell Biol 19:8314–8325
Lee S, Huang H, Niu Y et al (2007) Dok1 expression and mutation in Burkitt's lymphoma cell lines. Cancer Lett 245:44–50
Wu L, Bijian K, Shen SH (2009) CD45 recruits adapter protein DOK-1 and negatively regulates JAK-STAT signaling in hematopoietic cells. Mol Immunol 46:2167–2177
Cai D, Dhe-Paganon S, Melendez PA et al (2003) Two new substrates in insulin signaling, IRS5/DOK4 and IRS6/DOK5. J Biol Chem 278:25323–25330
Crowder RJ, Enomoto H, Yang M et al (2004) Dok-6, a Novel p62 Dok family member, promotes Ret-mediated neurite outgrowth. J Biol Chem 279:42072–42081
Grimm J, Sachs M, Britsch S et al (2001) Novel p62dok family members, dok-4 and dok-5, are substrates of the c-Ret receptor tyrosine kinase and mediate neuronal differentiation. J Cell Biol 154:345–354
Kurotsuchi A, Murakumo Y, Jijiwa M et al (2010) Analysis of DOK-6 function in downstream signaling of RET in human neuroblastoma cells. Cancer Sci 101:1147–1155
Hooker E, Baldwin C, Lemay S (2012) New insights into Dok-4 PTB domain structure and function. Biochem Biophys Res Commun 427:67–72
Niu Y, Roy F, Saltel F et al (2006) A nuclear export signal and phosphorylation regulate Dok1 subcellular localization and functions. Mol Cell Biol 26:4288–4301
Bhat A, Johnson KJ, Oda T et al (1998) Interactions of p62(dok) with p210(bcr-abl) and Bcr-Abl-associated proteins. J Biol Chem 273:32360–32368
Songyang Z, Yamanashi Y, Liu D et al (2001) Domain-dependent function of the rasGAP-binding protein p62Dok in cell signaling. J Biol Chem 276:2459–2465
Shinohara H, Yasuda T, Yamanashi Y (2004) Dok-1 tyrosine residues at 336 and 340 are essential for the negative regulation of Ras-Erk signalling, but dispensable for rasGAP-binding. Genes Cells 9:601–607
Zhao M, Janas JA, Niki M et al (2006) Dok-1 independently attenuates Ras/mitogen-activated protein kinase and Src/c-myc pathways to inhibit platelet-derived growth factor-induced mitogenesis. Mol Cell Biol 26:2479–2489
Downer EJ, Johnston DG, Lynch MA (2013) Differential role of Dok1 and Dok2 in TLR2-induced inflammatory signaling in glia. Mol Cell Neurosci 56:148–158
Oki S, Limnander A, Yao PM et al (2005) Dok1 and SHIP act as negative regulators of v-Abl-induced pre-B cell transformation, proliferation and Ras/Erk activation. Cell Cycle 4:310–314
Nold-Petry CA, Lo CY, Rudloff I et al (2015) IL-37 requires the receptors IL-18Rα and IL-1R8 (SIGIRR) to carry out its multifaceted anti-inflammatory program upon innate signal transduction. Nat Immunol 16:354–365
Yamakawa N, Tsuchida K, Sugino H (2002) The rasGAP-binding protein, Dok-1, mediates activin signaling via serine/threonine kinase receptors. EMBO J 21:1684–1694
Friedrich T, Söhn M, Gutting T et al (2016) Subcellular compartmentalization of docking protein-1 contributes to progression in colorectal cancer. EBioMedicine 8:159–172
Ghanem T, Bracken J, Kasem A et al (2014) mRNA expression of DOK1-6 in human breast cancer. World J Clin Oncol 5:156–163
Saulnier A, Vaissière T, Yue J et al (2012) Inactivation of the putative suppressor gene DOK1 by promoter hypermethylation in primary human cancers. Int J Cancer 130:2484–2494
He PF, Xu ZJ, Zhou JD et al (2018) Methylation-associated DOK1 and DOK2 down-regulation: potential biomarkers for predicting adverse prognosis in acute myeloid leukemia. J Cell Physiol 233(9):6604–6614
Araújo OC, Rosa AS, Fernandes A et al (2016) RASSF1A and DOK1 promoter methylation levels in hepatocellular carcinoma, cirrhotic and non-cirrhotic liver, and correlation with liver cancer in Brazilian patients. PLoS ONE 11:e0153796
Mercier PL, Bachvarova M, Plante M et al (2011) Characterization of DOK1, a candidate tumor suppressor gene, in epithelial ovarian cancer. Mol Oncol 5:438–453
Berger AH, Niki M, Morotti A et al (2010) Identification of DOK genes as lung tumor suppressors. Nat Genet 42:216–223
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
Ding X, Wang W, Wang M et al (2017) DOK1/PPARgamma pathway mediates anti-tumor ability of all-trans retinoic acid in breast cancer MCF-7 cells. Biochem Biophys Res Commun 487:189–193
Di Cristofano A, Niki M, Zhao M et al (2001) p62(dok), a negative regulator of Ras and mitogen-activated protein kinase (MAPK) activity, opposes leukemogenesis by p210(bcr-abl). J Exp Med 194:275–284
Lee S, Roy F, Galmarini CM et al (2004) Frameshift mutation in the Dok1 gene in chronic lymphocytic leukemia. Oncogene 23:2287–2297
Choi EJ, Lee JH, Kim MS et al (2018) Intratumoral heterogeneity of somatic mutations for NRIP1, DOK1, ULK1, ULK2, DLGAP3, PARD3 and PRKCI in colon cancers. Pathol Oncol Res 24(4):827–832
Conflict of interest
The authors declare that they have no conflict of interest.
The study was approved by the Cerrahpasa Medical Faculty Ethics Committee (Approval number: 7814), and has been performed in accordance with the ethical standarts laid down in the 2013 Declaration of Helsinki. Signed informed consent was obtained from all patients included in the study.
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Tuna, E., Ersoy, Y.E., Bulut, P. et al. Analysis of the DOK1 gene in breast cancer. Mol Biol Rep (2020). https://doi.org/10.1007/s11033-020-05247-3
- Breast cancer
- Mutation analysis