Knockdown of SET Domain, Bifurcated 1 suppresses head and neck cancer cell viability and wound-healing ability in vitro

Knockdown of SET Domain, Bifurcated 1 suppresses head and neck cancer cell viability and wound-healing ability in vitro

t: Head and neck cancer (HNC) is the sixth most common cancer worldwide and therefore presents a global public health problem. There are no standard algorithms for the diagnosis and follow-up of the disease, and no effective current treatment approachesexist. Therefore, the discovery of new biomolecules and the design of new strategies to aid in early diagnosis is necessary, along withestablishing prognostic factors of HNC. In several cancer studies, the upregulation of SET Domain, Bifurcated 1 (SETDB1) has beenreported to be tumor-inducing and to indicate a cancer-invasive prognosis, leading to the modulation of genes associated with differentsignaling pathways; however, the literature is sparse regarding the relationship between SETDB1 and HNC. In our study, three HNCprimary cell lines and their corresponding metastatic cell lines were used. The quantitative reverse transcriptase-polymerase chain reaction and western blotting data indicated that the SETDB1 mRNA and protein expression levels were higher in all metastatic cell linescompared to their primary cell lines (P < 0.05 for all). To investigate the role of SETDB1 in HNC biology, in vitro functional analyseswere carried out using small interference RNA (siRNA) technology, cell viability, scratch wound-healing, and the caspase-3 activity assay of gene expression of SETDB1 to compare primary and metastatic cell lines of HNC. Metastatic cells were more susceptible to thissuppression, which decreased the vitality of cells and their ability of wound-healing and induced level of caspase-3 activity (P < 0.05 forall). This functional study has shown that SETDB1 plays an important role in head and neck carcinogenesis. Therefore, SETDB1 may bean attractive therapeutic target molecule and also a potential diagnostic and prognostic biomarker in HNC.

___

  • Bakhtiar SM, Ali A, Barh D (2015). Epigenetics in head and neck cancer. Methods Molecular Biology 1238: 751-769.
  • Baylin SB, Ohm JE (2006). Epigenetic gene silencing in cancer - a mechanism for early oncogenic pathway addiction? Nature Reviews Cancer 6: 107-116.
  • Castilho RM, Squarize CH, Almeida LO (2017). Epigenetic modifications and head and neck cancer: implications for tumor progression and resistance to therapy. International Journal of Molecular Sciences 18 (7): E1506.
  • Ceol CJ, Houvras Y, Jane-Valbuena J, Bilodeau S, Orlando DA et al. (2011). The histone methyltransferase SETDB1 is recurrently amplified in melanoma and accelerates its onset. Nature 471: 513-517.
  • Chen K, Zhang F, Ding J, Liang Y, Zhan Z et al. (2017). Histone methyltransferase SETDB1 promotes the progression of colorectal cancer by inhibiting the expression of TP53. Journal of Cancer 16; 8 (16): 3318-3330.
  • Chiba T, Saito T, Yuki K, Zen Y, Koide S et al. (2015). Histone lysine methyltransferase SUV39H1 is a potent target for epigenetic therapy of hepatocellular carcinoma. International Journal of Cancer 15 136 (2): 289-298.
  • Dawson MA, Kouzarides T (2012). Cancer epigenetics: from mechanism to therapy. Cell 150: 12-27.
  • Clause KC, Barker TH (2013). Extracellular matrix signaling in morphogenesis and repair. Current Opinion in Biotechnology 24: 830-833.
  • Dodge JE, Kang YK, Beppu H, Lei H, Li E (2004). Histone H3-K9 methyltransferase ESET is essential for early development. Molecular and Cellular Biology 24: 2478-2486.
  • Fritsch L, Robin P, Mathieu JR, Souidi M, Hinaux H et al. (2010). A subset of the histone H3 lysine 9 methyltransferases Suv39h1, G9a, GLP, and SETDB1 participate in a multimeric complex. Molecular Cell 37 (1): 46-56.
  • Hua KT, Wang MY, Chen MW, Wei LH, Chen CK et al. (2014). The H3K9 methyltransferase G9a is a marker of aggressive ovarian cancer that promotes peritoneal metastasis. Molecular Cancer 13: 189.
  • Huang J, Huang W, Liu M, Zhu J, Jiang D et al. (2018). Enhanced expression of SETDB1 possesses prognostic value and promotes cell proliferation, migration and invasion in nasopharyngeal carcinoma. Oncology Reports 40 (2): 1017-1025.
  • Jemal A, Siegel R, Ward E, Murray T, Xu J et al. (2007). Cancer statistics. CA: A Cancer Journal for Clinicians 57 (1): 43-66.
  • Karanth AV, Maniswami RR, Prashanth S, Govindaraj H, Padmavathy R et al. (2017). Emerging role of SETDB1 as a therapeutic target. Expert Opinion on Therapeutic Targets 21 (3): 319-331.
  • Ko SK, Kim SK, Share A, Lynch VM, Park J et al. (2014). Synthetic ion transporters can induce apoptosis by facilitating chloride anion transport into cells. Nature Chemistry 6 (10): 885-892.
  • Kostaki M, Manona AD, Stavraka I, Korkolopoulou P, Levidou G et al. (2014). High-frequency p16(INK) (4A) promoter methylation is associated with histone methyltransferase SETDB1 expression in sporadic cutaneous melanoma. Experimental Dermatology 23: 332-338.
  • Lin CJ, Grandis JR, Carey TE, Gollin SM, Whiteside TL et al. (2007). Head and neck squamous cell carcinoma cell lines: established models and rationale for selection. Head & Neck 29: 163-188.
  • Lindgren D, Sjödahl G, Lauss M, Staaf J, Chebil G et al. (2012). Integrated genomic and gene expression profiling identifies two major genomic circuits in urothelial carcinoma. PLoS One 7: e38863.
  • Liu L, Kimball S, Liu H, Holowatyj A, Yang ZQ (2015). Genetic alterations of histone lysine methyltransferases and their significance in breast cancer. Oncotarget 6: 2466-2482.
  • Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25 (4): 402-408.
  • Matsui T, Leung D, Miyashita H, Maksakova IA, Miyachi H et al. (2010). Proviral silencing in embryonic stem cells requires the histone methyltransferase ESET. Nature 464 (7290): 927-931.
  • Matsui T, Leung D, Miyashita H, Maksakova IA, Miyachi H et al. (2010). Proviral silencing in embryonic stem cells requires the histone methyltransferase ESET. Nature 464: 927-931.
  • Minkovsky A, Sahakyan A, Rankin-Gee E, Bonora G, Patel S (2014). The Mbd1-Atf7ip-Setdb1 pathway contributes to the maintenance of X chromosome inactivation. Epigenetics Chromatin 7: 12.
  • Mochizuki D, Misawa Y, Kawasaki H, Imai A, Endo S et al. (2018). Aberrant epigenetic regulation in head and neck cancer due to distinct EZH2 overexpression and DNA hypermethylation. International Journal of Molecular Sciences 19 (12): E3707.
  • Na HH, Noh HJ, Cheong HM, Kang Y, Kim KC (2016). SETDB1 mediated FosB expression increases the cell proliferation rate during anticancer drug therapy. BMB Reports 49: 238-243.
  • Özdaş S (2018). Nuclear entrapment of p33ING1b by inhibition of exportin-1: A trigger of apoptosis in head and neck squamous cell cancer. Cellular and Molecular Biology (Noisy le Grand) 64: 66-72.
  • Özdaş S, Özdaş T (2018). Crm1-knockdown by specific small interfering RNA reduces cell proliferation and induces apoptosis in head and neck cancer cell lines. Turkish Journal of Biology 42: 132-143.
  • Pehlivan D, Gunduz E, Gunduz M, Nagatsuka H, Beder LB et al. (2008). Loss of heterozygosity at chromosome 14q is associated with poor prognosis in head and neck squamous cell carcinomas. Journal of Cancer Research and Clinical Oncology 134 (12): 1267-1276.
  • Porter AG, Jänicke RU (1999). Emerging roles of caspase-3 in apoptosis. Cell Death & Differentiation 6 (2): 99-104. Renan MJ (1993). How many mutations are required for tumorigenesis? Implications from human cancer data. Molecular Carcinogenesis 7: 139-146.
  • Richter GH, Plehm S, Fasan A, Rossler S, Unland R et al. (2009). EZH2 is a mediator of EWS/FLI1 driven tumor growth and metastasis blocking endothelial and neuro-ectodermal differentiation. Proceedings of the National Academy of Sciences of the United States of America 106: 5324-5329.
  • Rodriguez-Paredes M, Martinez de Paz A, Simó-Riudalbas L, Sayols S, Moutinho C et al. (2014). Gene amplification of the histone methyltransferase SETDB1 contributes to human lung tumorigenesis. Oncogene 33: 2807-2813.
  • Rorth P (2012). Fellow travellers: emergent properties of collective cell migration EMBO Reports 13: 984-991.
  • Ryu TY, Kim K, Kim SK, Oh JH, Min JK et al. (2019). SETDB1 regulates SMAD7 expression for breast cancer metastasis. BMB Reports 52: 139-144.
  • Taylor SC, Berkelman T, Yadav G, Hammond M (2013). A defined methodology for reliable quantification of Western blot data. Molecular Biotechnology 55 (3): 217-226.
  • Schultz DC, Ayyanathan K, Negorev D, Maul GG, Rauscher FJ (2002). SETDB1 a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins. Genes & Development 16: 919-932.
  • Shinoda N, Obata F, Zhang L, Miura M (2016). Drosophila SETDB1 and caspase cooperatively fine-tune cell fate determination of sensory organ precursor. Genes to Cells 21 (4): 378-386.
  • Song YJ, Choi JH, Lee H (2015). Setdb1 is required for myogenic differentiation of C2C12 myoblast cells via maintenance of MyoD expression. Molecular Cells 38: 362-372.
  • Spyropoulou A, Gargalionis A, Dalagiorgou G, Adamopoulos C, Papavassiliou KA et al. (2014). Role of histone lysine methyltransferases SUV39H1 and SETDB1 in gliomagenesis: modulation of cell proliferation, migration, and colony formation. Neuromolecular Medicine 16: 70-82.
  • Sun QY, Ding LW, Xiao JF, Chien W, Lim SL et al. (2015). SETDB1 accelerates tumourigenesis by regulating the WNT signalling pathway. Journal of Pathology 235 (4): 559-570.
  • Sun Y, Wei M, Ren SC, Chen R, Xu WD et al. (2014). Histone methyltransferase SETDB1 is required for prostate cancer cell proliferation, migration and invasion. Asian Journal of Andrology 16: 319-324.
  • Van Dyke DL, Worsham MJ, Benninger MS, Krause CJ, Baker SR et al. (1994). Recurrent cytogenetic abnormalities in squamous cell carcinomas of the head and neck region. Genes, Chromosomes and Cancer 9 (3): 192-206.
  • Wang H, An W, Cao R, Xia L, Erdjument-Bromage H et al. (2003). mAM facilitates conversion by ESET of dimethyl to trimethyl lysine 9 of histone H3 to cause transcriptional repression. Molecular Cell 12 (2): 475-487.
  • Wong CM, Wei L, Law CT, Ho DW, Tsang FH et al. (2016). Upregulation of histone methyltransferase SETDB1 by multiple mechanisms in hepatocellular carcinoma promotes cancer metastasis. Hepatology 63: 474-487.
  • Wu PC, Lu JW, Yang JY, Lin IH, Ou DL et al. (2014). H3K9 histone methyltransferase, KMT1E/SETDB1, cooperates with the SMAD2/3 pathway to suppress lung cancer metastasis. Cancer Research 74: 7333-7343.
  • Wu X, Yang L, Zheng Z, Li Z, Shi J et al (2016). Src promotes cutaneous wound healing by regulating MMP2 through the ERK pathway. International Journal of Molecular Medicine 37 (3): 639-648.
  • Zeng J, Ge Z, Wang L, Li Q, Wang N et al. (2010). The histone demethylase RBP2 is overexpressed in gastric cancer and its inhibition triggers senescence of cancer cells. Gastroenterology 138: 981-992.
  • Zhao M, Sano D, Pickering CR, Jasser SA, Henderson YC et al. (2011). Assembly and initial characterization of a panel of 85 genomically validated cell lines from diverse head and neck tumor sites. Clinical Cancer Research 17 (23): 7248-7264.