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Knockdown of zinc finger protein, X-linked (ZFX) inhibits cell proliferation and induces apoptosis in human laryngeal squamous cell carcinoma

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Abstract

ZFX (zinc finger protein, X-linked) gene locus on the human X chromosome is structurally similar to the zinc finger protein, Y-linked gene, which may constitute the primary sex-determining signal. However, the pathological roles of the dysfunction of ZFX gene in human disease such as cancer have not been addressed. Here, we analyzed the expression of ZFX in human laryngeal squamous cell carcinoma (LSCC) tissue specimens and found a significant up-regulation compared to corresponding non-tumorous LSCC tissue. Recombinant lentivirus expressing ZFX short hairpin RNA (shZFX) was constructed and infected Hep-2 human LSCC cells. We found that knockdown of ZFX gene resulted in suppression of proliferation and colony-forming ability of Hep-2 cells, and led to S phase cell cycle arrest. In addition, down-regulation of ZFX induced a significant enhancement of cell apoptosis and expression changes of apoptosis-related genes. These results suggest that high expression of ZFX is associated with LSCC progression and knockdown of ZFX may block tumor cell growth mainly by promoting cell apoptosis.

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References

  1. Parkin DM, Bray F, Ferlay J, Pisani P (2005) Global cancer statistics, 2002. CA Cancer J Clin 55(2):74–108

    Article  PubMed  Google Scholar 

  2. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ (2008) Cancer statistics, 2008. CA Cancer J Clin 58(2):71–96. doi:10.3322/CA.2007.0010

    Article  PubMed  Google Scholar 

  3. Marur S, Forastiere AA (2008) Head and neck cancer: changing epidemiology, diagnosis, and treatment. Mayo Clin Proc 83(4):489–501

    Article  PubMed  Google Scholar 

  4. Schnabl B, Valletta D, Kirovski G, Hellerbrand C (2011) Zinc finger protein 267 is up-regulated in hepatocellular carcinoma and promotes tumor cell proliferation and migration. Exp Mol Pathol. doi:10.1016/j.yexmp.2011.07.006

  5. Hu D, Wan Y (2011) Regulation of Kruppel-like factor 4 by the anaphase promoting complex pathway is involved in TGF-beta signaling. J Biol Chem 286(9):6890–6901. doi:10.1074/jbc.M110.179952

    Article  PubMed  CAS  Google Scholar 

  6. Cobaleda C, Perez-Caro M, Vicente-Duenas C, Sanchez-Garcia I (2007) Function of the zinc-finger transcription factor SNAI2 in cancer and development. Annu Rev Genet 41:41–61. doi:10.1146/annurev.genet.41.110306.130146

    Article  PubMed  CAS  Google Scholar 

  7. Zimmerer EJ, Threlkeld L (1995) A ZFY-like sequence in fish, with comments on the evolution of the ZFY family of genes in vertebrates. Biochem Genet 33(7–8):227–235

    PubMed  CAS  Google Scholar 

  8. North M, Sargent C, O’Brien J, Taylor K, Wolfe J, Affara NA, Ferguson-Smith MA (1991) Comparison of ZFY and ZFX gene structure and analysis of alternative 3′ untranslated regions of ZFY. Nucleic Acids Res 19(10):2579–2586

    Article  PubMed  CAS  Google Scholar 

  9. Tucker PK, Adkins RM, Rest JS (2003) Differential rates of evolution for the ZFY-related zinc finger genes, Zfy, Zfx, and Zfa in the mouse genus Mus. Mol Biol Evol 20(6):999–1005. doi:10.1093/molbev/msg112

    Article  PubMed  CAS  Google Scholar 

  10. Schneider-Gadicke A, Beer-Romero P, Brown LG, Nussbaum R, Page DC (1989) ZFX has a gene structure similar to ZFY, the putative human sex determinant, and escapes X inactivation. Cell 57(7):1247–1258

    Article  PubMed  CAS  Google Scholar 

  11. Ramalho-Santos M, Yoon S, Matsuzaki Y, Mulligan RC, Melton DA (2002) “Stemness”: transcriptional profiling of embryonic and adult stem cells. Science 298(5593):597–600. doi:10.1126/science.1072530

    Article  PubMed  CAS  Google Scholar 

  12. Galan-Caridad JM, Harel S, Arenzana TL, Hou ZE, Doetsch FK, Mirny LA, Reizis B (2007) Zfx controls the self-renewal of embryonic and hematopoietic stem cells. Cell 129(2):345–357. doi:10.1016/j.cell.2007.03.014

    Article  PubMed  CAS  Google Scholar 

  13. Arenzana TL, Smith-Raska MR, Reizis B (2009) Transcription factor Zfx controls BCR-induced proliferation and survival of B lymphocytes. Blood 113(23):5857–5867. doi:10.1182/blood-2008-11-188888

    Article  PubMed  CAS  Google Scholar 

  14. Soneoka Y, Cannon PM, Ramsdale EE, Griffiths JC, Romano G, Kingsman SM, Kingsman AJ (1995) A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res 23(4):628–633

    Article  PubMed  CAS  Google Scholar 

  15. Sakoda T, Kasahara N, Hamamori Y, Kedes L (1999) A high-titer lentiviral production system mediates efficient transduction of differentiated cells including beating cardiac myocytes. J Mol Cell Cardiol 31(11):2037–2047. doi:10.1006/jmcc.1999.1035

    Article  PubMed  CAS  Google Scholar 

  16. Tiscornia G, Singer O, Verma IM (2006) Production and purification of lentiviral vectors. Nat Protoc 1(1):241–245. doi:10.1038/nprot.2006.37

    Article  PubMed  CAS  Google Scholar 

  17. 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(4):402–408. doi:10.1006/meth.2001.1262

    Article  PubMed  CAS  Google Scholar 

  18. Tricoli JV, Bracken RB (1993) ZFY gene expression and retention in human prostate adenocarcinoma. Genes Chromosomes Cancer 6(2):65–72

    Article  PubMed  CAS  Google Scholar 

  19. Kim DH, Behlke MA, Rose SD, Chang MS, Choi S, Rossi JJ (2005) Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy. Nat Biotechnol 23(2):222–226. doi:10.1038/nbt1051

    Article  PubMed  CAS  Google Scholar 

  20. Manilla P, Rebello T, Afable C, Lu X, Slepushkin V, Humeau LM, Schonely K, Ni Y, Binder GK, Levine BL, MacGregor RR, June CH, Dropulic B (2005) Regulatory considerations for novel gene therapy products: a review of the process leading to the first clinical lentiviral vector. Hum Gene Ther 16(1):17–25. doi:10.1089/hum.2005.16.17

    Article  PubMed  CAS  Google Scholar 

  21. Bank A, Dorazio R, Leboulch P (2005) A phase I/II clinical trial of beta-globin gene therapy for beta-thalassemia. Ann N Y Acad Sci 1054:308–316. doi:10.1196/annals.1345.007

    Article  PubMed  CAS  Google Scholar 

  22. Black AR, Black JD, Azizkhan-Clifford J (2001) Sp1 and Kruppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol 188(2):143–160. doi:10.1002/jcp.1111

    Article  PubMed  CAS  Google Scholar 

  23. Hunter AM, LaCasse EC, Korneluk RG (2007) The inhibitors of apoptosis (IAPs) as cancer targets. Apoptosis 12(9):1543–1568. doi:10.1007/s10495-007-0087-3

    Article  PubMed  CAS  Google Scholar 

  24. Nam SY, Seo HH, Park HS, An S, Kim JY, Yang KH, Kim CS, Jeong M, Jin YW (2010) Phosphorylation of CLK2 at serine 34 and threonine 127 by AKT controls cell survival after ionizing radiation. J Biol Chem 285(41):31157–31163. doi:10.1074/jbc.M110.122044

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

The authors are thankful for the financial support from National Natural Science Foundation of China (81072204).

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Authors and Affiliations

  1. Department of Otolaryngology, Head and Neck Surgery, Beijing TongRen Hospital, Capital Medical University, No. 1 Dong Jiao Min Xiang, Eastern District, Beijing, 100730, People’s Republic of China

    Jugao Fang, Zhenkun Yu, Meng Lian, Hongzhi Ma, Jun Tai, Luo Zhang & Demin Han

  2. Key Laboratory of Otolaryngology, Head and Neck Surgery (Ministry of Education of China), Beijing Institute of Otorhinolaryngology, No. 17 Hou Gou Lane, Chong Nei Street, Beijing, 100005, People’s Republic of China

    Jugao Fang, Zhenkun Yu, Meng Lian, Hongzhi Ma, Jun Tai, Luo Zhang & Demin Han

Authors
  1. Jugao Fang
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  2. Zhenkun Yu
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  3. Meng Lian
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  4. Hongzhi Ma
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  5. Jun Tai
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  6. Luo Zhang
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  7. Demin Han
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Corresponding authors

Correspondence to Luo Zhang or Demin Han.

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Demin Han and Luo Zhang are contributed equally to this study.

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11010_2011_1069_MOESM1_ESM.tif

Effect of pGCSIL-GFP-shZFX′ lentivirus infection on Hep-2 cell growth. a Cell count assay. ZFX knockdown suppressed the growth of Hep-2 cells. b Colony formation assay. ZFX knockdown inhibited colony-forming capacity of Hep-2 cells. c Analysis of cell cycle by flow cytometry. ZFX knockdown induced cell cycle arrest at S-phase. d Flow cytometric analysis of Annexin V-APC staining. ZFX knockdown induced Hep-2 cell apoptosis. The value represents the mean from three independent experiments; bars, SD. *P < 0.05, **P < 0.01, ***P < 0.001, in comparison with control. (TIFF 204 kb)

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Fang, J., Yu, Z., Lian, M. et al. Knockdown of zinc finger protein, X-linked (ZFX) inhibits cell proliferation and induces apoptosis in human laryngeal squamous cell carcinoma. Mol Cell Biochem 360, 301–307 (2012). https://doi.org/10.1007/s11010-011-1069-x

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  • DOI: https://doi.org/10.1007/s11010-011-1069-x

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