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Tumor Biology

, Volume 35, Issue 6, pp 5219–5225 | Cite as

Mutations and Krüppel-like factor 6 (KLF6) expression levels in breast cancer

  • Filiz Ozdemir
  • Mehtap Koksal
  • Vahit Ozmen
  • Ibrahim Aydin
  • Nur BuyruEmail author
Research Article
First Online:

Abstract

The transcription factor KLF6 gene has been identified as a tumor suppressor because of its inactivation in several types of cancers by different mechanisms. However, there are no data in the literature investigating the KLF6 mutation rate and expression levels in breast cancer. Therefore, the present study was conducted in order to investigate whether genetic alterations of KLF6 in association with the KLF6 mRNA expression levels may play a role in breast carcinogenesis. For this purpose, we analyzed alterations of the KLF6 gene by direct sequencing and the mRNA levels by reverse transcription-PCR (RT-PCR). In addition to four different non-coding alterations, one missense and two silent alterations were identified in the coding sequence. Reduced KLF6 expression was observed in 41 (83.67 %) of the 49 breast cancer tumors. These findings suggest that the mutation profile of the KLF6 gene in breast tumors is similar to other cancer types. However, these mutations do not exert any effect on the gene expression rate. Downregulation of KLF6 during the progression of breast cancer is independent of the mutations and occurs by a different mechanism.

Keywords

Breast cancer KLF6 Expression Real-time PCR Tumor suppressor gene 
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Notes

Acknowledgments

The present work was supported by the Research Fund of Istanbul University (Project No: 3439 ).

Conflict of interest

None

References

  1. 1.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90. doi: 10.3322/caac.20107.CrossRefPubMedGoogle Scholar
  2. 2.
    Bieker JJ. Krüppel-like factors: three fingers in many pies. J Biol Chem. 2001;276:34355–8.CrossRefPubMedGoogle Scholar
  3. 3.
    Black AR, Black JD, Azizkhan-Clifford J. Sp1 and krüppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol. 2001;188:143–60.CrossRefPubMedGoogle Scholar
  4. 4.
    Pei J, Grishin NV. A new family of predicted Krüppel-like factor genes and pseudogenes in placental mammals. PLoS One. 2013;8(11):e81109. doi: 10.1371/journal.pone.0081109.PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Schuh R, Aicher W, Gaul U, Côté S, Preiss A, Maier D, et al. A conserved family of nuclear proteins containing structural elements of the finger protein encoded by Krüppel, a Drosophila segmentation gene. Cell. 1986;47:1025–32.CrossRefPubMedGoogle Scholar
  6. 6.
    Pearson R, Fleetwood J, Eaton S, Crossley M, Bao S. Krüppel-like transcription factors: a functional family. Int J Biochem Cell Biol. 2008;40:1996–2001.CrossRefPubMedGoogle Scholar
  7. 7.
    Narla G, Heath KE, Reeves HL, Li D, Giono LE, Kimmelman AC, et al. KLF6, a candidate tumor suppressor gene mutated in prostate cancer. Science. 2001;294:2563–6.CrossRefPubMedGoogle Scholar
  8. 8.
    Ittmann M. Allelic loss on chromosome 10 in prostate adenocarcinoma. Cancer Res. 1996;56:2143–7.PubMedGoogle Scholar
  9. 9.
    Koritschoner NP, Bocco JL, Panzetta-Dutari GM, Dumur CI, Flury A, Patrito LC. A novel human zinc finger protein that interacts with the core promoter element of a TATA box-less gene. J Biol Chem. 1997;272:9573–80.CrossRefPubMedGoogle Scholar
  10. 10.
    Miyaki M, Yamaguchi T, Iijima T, Funata N, Mori T. Difference in the role of loss of heterozygosity at 10p15 (KLF6 locus) in colorectal carcinogenesis between sporadic and familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer patients. Oncology. 2006;71:131–5.CrossRefPubMedGoogle Scholar
  11. 11.
    Mukai S, Hiyama T, Tanaka S, Yoshihara M, Arihiro K, Chayama K. Involvement of Kruppel-like factor 6 (KLF6) mutation in the development of nonpolypoid colorectal carcinoma. World J Gastroenterol. 2007;13:3932–8.PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Ito G, Uchiyama M, Kondo M, Mori S, Usami N, Maeda O, et al. Krüppel-like factor 6 is frequently down-regulated and induces apoptosis in non-small cell lung cancer cells. Cancer Res. 2004;64:3838–43.CrossRefPubMedGoogle Scholar
  13. 13.
    Sangodkar J, Shi J, DiFeo A, Schwartz R, Bromberg R, Choudhri A, et al. Functional role of the KLF6 tumour suppressor gene in gastric cancer. Eur J Cancer. 2009;45:666–76. doi: 10.1016/j.ejca.2008.11.009.PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Camacho-Vanegas O, Narla G, Teixeira MS, DiFeo A, Misra A, Singh G, et al. Functional inactivation of the KLF6 tumor suppressor gene by loss of heterozygosity and increased alternative splicing in glioblastoma. Int J Cancer. 2007;121:1390–5.CrossRefPubMedGoogle Scholar
  15. 15.
    Teixeira MS, Camacho-Vanegas O, Fernandez Y, Narla G, DiFeo A, Lee B, et al. KLF6 allelic loss is associated with tumor recurrence and markedly decreased survival in head and neck squamous cell carcinoma. Int J Cancer. 2007;121:1976–83.CrossRefPubMedGoogle Scholar
  16. 16.
    Song J, Kim CJ, Cho YG, Kim SY, Nam SW, Lee SH, et al. Genetic and epigenetic alterations of the KLF6 gene in hepatocellular carcinoma. J Gastroenterol Hepatol. 2006;21:1286–9.CrossRefPubMedGoogle Scholar
  17. 17.
    Yea S, Narla G, Zhao X, Garg R, Tal-Kremer S, Hod E, et al. Ras promotes growth by alternative splicing-mediated inactivation of the KLF6 tumor suppressor in hepatocellular carcinoma. Gastroenterology. 2008;134:1521–31. doi: 10.1053/j.gastro.2008.02.015.PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    DiFeo A, Narla G, Hirshfeld J, Camacho-Vanegas O, Narla J, Rose SL, et al. Roles of KLF6 and KLF6-SV1 in ovarian cancer progression and intraperitoneal dissemination. Clin Cancer Res. 2006;12:3730–9.CrossRefPubMedGoogle Scholar
  19. 19.
    Benzeno S, Narla G, Allina J, Cheng GZ, Reeves HL, Banck MS, et al. Cyclin-dependent kinase inhibition by the KLF6 tumor suppressor protein through interaction with cyclin D1. Cancer Res. 2004;64:3885–91.CrossRefPubMedGoogle Scholar
  20. 20.
    Sirach E, Bureau C, Péron JM, Pradayrol L, Vinel JP, Buscail L, et al. KLF6 transcription factor protects hepatocellular carcinoma-derived cells from apoptosis. Cell Death Differ. 2007;14:1202–10.CrossRefPubMedGoogle Scholar
  21. 21.
    Slavin DA, Koritschoner NP, Prieto CC, López-Díaz FJ, Chatton B, Bocco JL. A new role for the Krüppel-like transcription factor KLF6 as an inhibitor of c-Jun proto-oncoprotein function. Oncogene. 2004;23:8196–205.CrossRefPubMedGoogle Scholar
  22. 22.
    Gehrau RC, D'Astolfo DS, Dumur CI, Bocco JL, Koritschoner NP. Nuclear expression of KLF6 tumor suppressor factor is highly associated with overexpression of ERBB2 oncoprotein in ductal breast carcinomas. PLoS One. 2010;5:e8929. doi: 10.1371/journal.pone.0008929.PubMedCentralCrossRefPubMedGoogle Scholar
  23. 23.
    Liu J, Du T, Yuan Y, He Y, Tan Z, Liu Z. KLF6 inhibits estrogen receptor-mediated cell growth in breast cancer via a c-Src-mediated pathway. Mol Cell Biochem. 2010;335:29–35. doi: 10.1007/s11010-009-0237-8.CrossRefPubMedGoogle Scholar
  24. 24.
    Sommer S, Fuqua SA. Estrogen receptor and breast cancer. Semin Cancer Biol. 2001;11:339–52.CrossRefPubMedGoogle Scholar
  25. 25.
    Narla G, Difeo A, Reeves HL, Schaid DJ, Hirshfeld J, Hod E, et al. A germline polymorphism enhances alternative splicing of the KLF6 tumor suppressor gene and is associated with increased prostate cancer risk. Cancer Res. 2005;65:1213–22.CrossRefPubMedGoogle Scholar
  26. 26.
    DiFeo A, Feld L, Rodriguez E, Wang C, Beer DG, Martignetti JA, et al. A functional role for KLF6-SV1 in lung adenocarcinoma prognosis and chemotherapy response. Cancer Res. 2008;68:965–70. doi: 10.1158/0008-5472.CAN-07-2604.PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Agell L, Hernandez S, de Muga S, Lorente JA, Juanpere N, Esgueva R, et al. KLF6 and TP53 mutations are rare event in prostate cancer: distinguishing between Taq polymerase artifacts and true mutations. Mod Pathol. 2008;21:1470–8. doi: 10.1038/modpathol.2008.145.CrossRefPubMedGoogle Scholar
  28. 28.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001;25:402–8.CrossRefPubMedGoogle Scholar
  29. 29.
    Chen C, Hyytinen ER, Sun X, Helin HJ, Koivisto PA, Frierson Jr HF, et al. Deletion, mutation, and loss of expression of KLF6 in human prostate cancer. Am J Pathol. 2003;162:1349–54.PubMedCentralCrossRefPubMedGoogle Scholar
  30. 30.
    Huang X, Li X, Guo B. KLF6 induces apoptosis in prostate cancer cells through up-regulation of ATF3. J Biol Chem. 2008;283:29795–801. doi: 10.1074/jbc.M802515200.PubMedCentralCrossRefPubMedGoogle Scholar
  31. 31.
    Narla G, Friedman SL, Martignetti JA. Krüppel cripples prostate cancer: KLF6 progress and prospects. Am J Pathol. 2003;162:1047–52.PubMedCentralCrossRefPubMedGoogle Scholar
  32. 32.
    Chen HK, Liu XQ, Lin J, Chen TY, Feng QS, Zeng YX. Mutation analysis of KLF6 gene in human nasopharyngeal carcinomas. Ai Zheng. 2002;21:1047–50.PubMedGoogle Scholar
  33. 33.
    Jeng YM, Hsu HC. KLF6, a putative tumor suppressor gene, is mutated in astrocytic gliomas. Int J Cancer. 2003;105:625–9.CrossRefPubMedGoogle Scholar
  34. 34.
    Koivisto PA, Hyytinen ER, Matikainen M, Tammela TL, Ikonen T, Schleutker J. Kruppel-like factor 6 germ-line mutations are infrequent in Finnish hereditary prostate cancer. J Urol. 2004;172:506–7.CrossRefPubMedGoogle Scholar
  35. 35.
    Boyault S, Hérault A, Balabaud C, Zucman-Rossi J. Absence of KLF6 gene mutation in 71 hepatocellular carcinomas. Hepatology. 2005;41:681–2.CrossRefPubMedGoogle Scholar
  36. 36.
    Lièvre A, Landi B, Côté JF, Veyrie N, Zucman-Rossi J, Berger A, et al. Absence of mutation in the putative tumor-suppressor gene KLF6 in colorectal cancers. Oncogene. 2005;24:7253–6.CrossRefPubMedGoogle Scholar
  37. 37.
    Yin D, Komatsu N, Miller CW, Chumakov AM, Marschesky A, McKenna R, et al. KLF6: mutational analysis and effect on cancer cell proliferation. Int J Oncol. 2007;30:65–72.PubMedGoogle Scholar
  38. 38.
    Vax VV, Gueorguiev M, Dedov II, Grossman AB, Korbonits M. The Krüppel-like transcription factor 6 gene in sporadic pituitary tumours. Endocr Relat Cancer. 2003;10:397–402.CrossRefPubMedGoogle Scholar
  39. 39.
    Pandya AY, Talley LI, Frost AR, Fitzgerald TJ, Trivedi V, Chakravarthy M, et al. Nuclear localization of KLF4 is associated with an aggressive phenotype in early-stage breast cancer. Clin Cancer Res. 2004;10:2709–19.CrossRefPubMedGoogle Scholar
  40. 40.
    Chen C, Bhalala HV, Qiao H, Dong JT. A possible tumor suppressor role of the KLF5 transcription factor in human breast cancer. Oncogene. 2002;21:6567–72.CrossRefPubMedGoogle Scholar
  41. 41.
    Hatami R, Sieuwerts AM, Izadmehr S, Yao Z, Qiao RF, Papa L, et al. KLF6-SV1 drives breast cancer metastasis and is associated with poor survival. Sci Transl. Med. 2013; 5:169ra12. doi:  10.1126/scitranslmed.3004688.
  42. 42.
    Narla G, DiFeo A, Yao S, Banno A, Hod E, Reeves HL, et al. Targeted inhibition of the KLF6 splice variant, KLF6 SV1, suppresses prostate cancer cell growth and spread. Cancer Res. 2005;65:5761–8.CrossRefPubMedGoogle Scholar
  43. 43.
    Narla G, DiFeo A, Fernandez Y, Dhanasekaran S, Huang F, Sangodkar J, et al. KLF-SV1 overexpression accelerates human and mouse prostate cancer progression and metastasis. J Clin Invest. 2008;118:2711–21. doi: 10.1172/JCI34780.PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Filiz Ozdemir
    • 1
  • Mehtap Koksal
    • 1
  • Vahit Ozmen
    • 2
  • Ibrahim Aydin
    • 3
  • Nur Buyru
    • 1
    Email author
  1. 1.Cerrahpasa Medical Faculty, Department of Medical BiologyIstanbul UniversityIstanbulTurkey
  2. 2.Istanbul Medical Faculty, Department of General SurgeryIstanbul UniversityIstanbulTurkey
  3. 3.Department of General SurgeryIstanbul Training and Research HospitalIstanbulTurkey

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