MicroRNA-196a-5p is a potential prognostic marker of delayed lymph node metastasis in early-stage tongue squamous cell carcinoma

Maruyama,Tessho; Nishihara,Kazuhide; Umikawa,Masato; Arasaki,Akira; Nakasone,Toshiyuki; Nimura,Fumikazu; Matayoshi,Akira; Takei,Kimiko; Nakachi,Saori; Kariya,Ken‑Ichi; Yoshimi,Naoki

doi:10.3892/ol.2017.7562

MicroRNA-196a-5p is a potential prognostic marker of delayed lymph node metastasis in early-stage tongue squamous cell carcinoma

Authors:
- Tessho Maruyama
- Kazuhide Nishihara
- Masato Umikawa
- Akira Arasaki
- Toshiyuki Nakasone
- Fumikazu Nimura
- Akira Matayoshi
- Kimiko Takei
- Saori Nakachi
- Ken‑Ichi Kariya
- Naoki Yoshimi
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Affiliations: Department of Oral and Maxillofacial Functional Rehabilitation, Graduate School of Medicine, University of the Ryukyus, Nishihara, Okinawa 903‑0215, Japan, Department of Medical Biochemistry, Graduate School of Medicine, University of the Ryukyus, Nishihara, Okinawa 903‑0215, Japan, Department of Oral and Maxillofacial Surgery, University Hospital of the Ryukyus, Nishihara, Okinawa 903‑0215, Japan, Department of Pathology, University Hospital of the Ryukyus, Nishihara, Okinawa 903‑0215, Japan
Published online on: December 8, 2017 https://doi.org/10.3892/ol.2017.7562
Pages: 2349-2363
Copyright: © Maruyama et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

MicroRNAs (miRs) are expected to serve as prognostic tools for cancer. However, many miRs have been reported as prognostic markers of recurrence or metastasis in oral squamous cell carcinoma patients. We aimed to determine the prognostic markers in early‑stage tongue squamous cell carcinoma (TSCC). Based on previous studies, we hypothesized that miR‑10a, 10b, 196a‑5p, 196a‑3p, and 196b were prognostic markers and we retrospectively performed miR expression analyses using formalin‑fixed paraffin‑embedded sections of surgical specimens. Total RNA was isolated from cancer tissues and adjacent normal tissue as control, and samples were collected by laser‑capture microdissection. After cDNA synthesis, reverse transcription‑quantitative polymerase chain reaction was performed. Statistical analyses for patient clinicopathological characteristics, recurrence/metastasis, and survival rates were performed to discern their relationships with miR expression levels, and the 2‑ΔΔCq method was used. miR‑196a‑5p levels were significantly upregulated in early‑stage TSCC, particularly in the lymph node metastasis (LNM) group. The LNM‑free survival rate in the low miR‑196a‑5p ΔΔCq value regulation group was found to be lower than that in the high ΔΔCq value regulation group (P=0.0079). Receiver operating characteristic analysis of ΔΔCq values revealed that miR‑196a‑5p had a P‑value=0.0025, area under the curve=0.740, and a cut‑off value=‑0.875 for distinguishing LNM. To our knowledge, this is the first study to examine LNM‑related miRs in early‑stage TSCC as well as miRs and ‘delayed LNM’ in head and neck cancer. miR‑196a‑5p upregulation may predict delayed LNM. Our data serve as a foundation for future studies to evaluate miR levels and facilitate the prediction of delayed LNM during early‑stage TSCC, which prevent metastasis when combined with close follow‑up and aggressive adjuvant therapy or elective neck dissection. Moreover, our data will serve as a foundation for future studies to evaluate whether miR‑196a‑5p can serve as a therapeutic marker for preventing metastasis.

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1	IARC: GLOBOCAN 2012: Estimated Cancer Incidence, Mortality and Prevalence Worldwide in 2012. http://globocan.iarc.fr/Pages/fact_sheets_population.aspxFebruary 22–2017
2	Brierley JD, Gospodarowicz M and Wittekind C: Lip and Oral CavityTNM Classification of Malignant Tumours. 8th edition. Wiley Blackwell; New York, NY: pp. 18–21. 2016
3	Sinevici N and O'sullivan J: Oral cancer: Deregulated molecular events and their use as biomarkers. Oral Oncol. 61:12–18. 2016. View Article : Google Scholar : PubMed/NCBI
4	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 660:7–30. 2016. View Article : Google Scholar
5	Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ and Thun MJ: Cancer statistics, 2005. CA Cancer J Clin. 55:2005. View Article : Google Scholar : PubMed/NCBI
6	Parker SL, Tong T, Bolden S and Wingo PA: Cancer statistics, 1996. CA Cancer J Clin. 46:5–27. 1996. View Article : Google Scholar : PubMed/NCBI
7	Li Z, Wang Y, Qiu J, Li Q, Yuan C, Zhang W, Wang D, Ye J, Jiang H, Yang J and Cheng J: The polycomb group protein EZH2 is a novel therapeutic target in tongue cancer. Oncotarget. 4:2532–2549. 2013. View Article : Google Scholar : PubMed/NCBI
8	Wang C, Huang H, Huang Z, Wang A, Chen X, Huang L, Zhou X and Liu X: Tumor budding correlates with poor prognosis and epithelial-mesenchymal transition in tongue squamous cell carcinoma. J Oral Pathol Med. 40:545–551. 2011. View Article : Google Scholar : PubMed/NCBI
9	Schwam ZG and Judson BL: Improved prognosis for patients with oral cavity squamous cell carcinoma: Analysis of the national cancer database 1998–2006. Oral Oncol. 52:45–51. 2016. View Article : Google Scholar : PubMed/NCBI
10	Yoon AJ, Wang S, Shen J, Robine N, Philipone E, Oster MW, Nam A and Santella RM: Prognostic value of miR-375 and miR-214-3p in early stage oral squamous cell carcinoma. Am J Transl Res. 6:580–592. 2014.PubMed/NCBI
11	National ComprehensiveCancer Network, . NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines): Head and Neck Cancers 2017. https://www.nccn.org/professionals/physician_gls/pdf/head-and-neck.pdfFebruary 24–2017
12	Rogers SN, Brown JS, Woolgar JA, Lowe D, Magennis P, Shaw RJ, Sutton D, Errington D and Vaughan D: Survival following primary surgery for oral cancer. Oral Oncol. 45:201–211. 2009. View Article : Google Scholar : PubMed/NCBI
13	Kelner N, Vartanian JG, Pinto CA, Coutinho-Camillo CM and Kowalski LP: Does elective neck dissection in T1/T2 carcinoma of the oral tongue and floor of the mouth influence recurrence and survival rates? Br J Oral Maxillofac Surg. 52:590–597. 2014. View Article : Google Scholar : PubMed/NCBI
14	Ong HS, Gokavarapu S, Wang LZ, Tian Z and Zhang CP: Low pretreatment lymphocyte-monocyte ratio and high platelet-lymphocyte ratio indicate poor cancer outcome in early tongue cancer. J Oral Maxillofac Surg. 75:1762–1774. 2017. View Article : Google Scholar : PubMed/NCBI
15	Yu X and Li Z: MicroRNA expression and its implications for diagnosis and therapy of tongue squamous cell carcinoma. J Cell Mol Med. 20:10–16. 2016. View Article : Google Scholar : PubMed/NCBI
16	Seki M, Sano T, Yokoo S and Oyama T: Tumour budding evaluated in biopsy specimens is a useful predictor of prognosis in patients with cN0 early stage oral squamous cell carcinoma. Histopathology. 70:869–879. 2017. View Article : Google Scholar : PubMed/NCBI
17	Ariji Y, Goto M, Fukano H, Sugita Y, Izumi M and Ariji E: Role of intraoral color Doppler sonography in predicting delayed cervical lymph node metastasis in patients with early-stage tongue cancer: A pilot study. Oral Surg Oral Med Oral Pathol Oral Radiol. 119:246–253. 2015. View Article : Google Scholar : PubMed/NCBI
18	Habu N, Imanishi Y, Kameyama K, et al: Expression of Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II oral tongue squamous cell carcinoma. BMC Cancer. 15:7302015. View Article : Google Scholar : PubMed/NCBI
19	Goto M, Hanai N, Ozawa T, Hirakawa H, Suzuki H, Hyodo I, Kodaira T, Ogawa T, Fujimoto Y, Terada A, et al: Prognostic factors and outcomes for salvage surgery in patients with recurrent squamous cell carcinoma of the tongue. Asia Pac J Clin Oncol. 12:e141–e148. 2016. View Article : Google Scholar : PubMed/NCBI
20	Luksic I and Suton P: Predictive markers for delayed lymph node metastases and survival in early-stage oral squamous cell carcinoma. Head Neck. 39:694–701. 2017. View Article : Google Scholar : PubMed/NCBI
21	Almangush A, Coletta RD, Bello IO, Bitu C, Keski-Säntti H, Mäkinen LK, Kauppila JH, Pukkila M, Hagström J, Laranne J, et al: A simple novel prognostic model for early stage oral tongue cancer. Int J Oral Maxillofac Surg. 44:143–150. 2015. View Article : Google Scholar : PubMed/NCBI
22	Montebugnoli L, Gissi DB, Flamminio F, Gentile L, Dallera V, Leonardi E, Beccarini T and Foschini MP: Clinicopathologic parameters related to recurrence and locoregional metastasis in 180 oral squamous cell carcinomas. Int J Surg Pathol. 22:55–62. 2014. View Article : Google Scholar : PubMed/NCBI
23	Dunkel J, Vaittinen S, Grénman R, Kinnunen I and Irjala H: Prognostic markers in stage I oral cavity squamous cell carcinoma. Laryngoscope. 123:2435–2441. 2013.PubMed/NCBI
24	Harada Y, Izumi H, Noguchi H, Kuma A, Kawatsu Y, Kimura T, Kitada S, Uramoto H, Wang KY, Sasaguri Y, et al: Erratum to: Strong expression of polypeptide N-acetylgalactosaminyltransferase 3 independently predicts shortened disease-free survival in patients with early stage oral squamous cell carcinoma. Tumour Biol. 36:10003–10004. 2015. View Article : Google Scholar : PubMed/NCBI
25	Villegas-Ruiz V, Juárez-Méndez S, Pérez-González OA, Arreola H, Paniagua-García L, Parra-Melquiadez M, Peralta-Rodríguez R, López-Romero R, Monroy-García A, Mantilla-Morales A, et al: Heterogeneity of microRNAs expression in cervical cancer cells: Over-expression of miR-196a. Int J Clin Exp Pathol. 7:1389–1401. 2014.PubMed/NCBI
26	Boldrup L, Coates PJ, Laurell G, Wilms T, Fahraeus R and Nylander K: Downregulation of miRNA-424: A sign of field cancerisation in clinically normal tongue adjacent to squamous cell carcinoma. Br J Cancer. 112:1760–1765. 2015. View Article : Google Scholar : PubMed/NCBI
27	Liu X, Wang A, Heidbreder CE, Jiang L, Yu J, Kolokythas A, Huang L, Dai Y and Zhou X: MicroRNA-24 targeting RNA-binding protein DND1 in tongue squamous cell carcinoma. FEBS Lett. 584:4115–4120. 2010. View Article : Google Scholar : PubMed/NCBI
28	Rosenfeld N, Aharonov R, Meiri E, Rosenwald S, Spector Y, Zepeniuk M, Benjamin H, Shabes N, Tabak S, Levy A, et al: MicroRNAs accurately identify cancer tissue origin. Nat Biotechnol. 26:462–469. 2008. View Article : Google Scholar : PubMed/NCBI
29	Liu CJ, Tsai MM, Tu HF, Lui MT, Cheng HW and Lin SC: miR-196a overexpression and miR-196a2 gene polymorphism are prognostic predictors of oral carcinomas. Ann Surg Oncol. Suppl. 3 20 Suppl:S406–S414. 2013.
30	Liu CJ, Lin SC, Yang CC, Cheng HW and Chang KW: Exploiting salivary miR-31 as a clinical biomarker of oral squamous cell carcinoma. Head Neck. 34:219–224. 2012. View Article : Google Scholar : PubMed/NCBI
31	Di Leva G, Garofalo M and Croce CM: MicroRNAs in cancer. Annu Rev Pathol. 9:287–314. 2014. View Article : Google Scholar : PubMed/NCBI
32	Reddy KB: MicroRNA (miRNA) in cancer. Cancer Cell Int. 15:382015. View Article : Google Scholar : PubMed/NCBI
33	Lin S and Gregory RI: MicroRNA biogenesis pathways in cancer. Nat Rev Cancer. 15:321–333. 2015. View Article : Google Scholar : PubMed/NCBI
34	Lu YC, Chang JT, Chan EC, Chao YK, Yeh TS, Chen JS and Cheng AJ: miR-196, an emerging cancer biomarker for digestive tract cancers. J Cancer. 7:650–655. 2016. View Article : Google Scholar : PubMed/NCBI
35	Chen ZY, Chen X and Wang ZX: The role of microRNA-196a in tumorigenesis, tumor progression, and prognosis. Tumour Biol. Oct 18–2016.(Epub ahead of print). View Article : Google Scholar
36	Friedman RC, Farh KK, Burge CB and Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19:92–105. 2009. View Article : Google Scholar : PubMed/NCBI
37	Nohata N, Hanazawa T, Kinoshita T, Okamoto Y and Seki N: MicroRNAs function as tumor suppressors or oncogenes: Aberrant expression of microRNAs in head and neck squamous cell carcinoma. Auris Nasus Larynx. 40:143–149. 2013. View Article : Google Scholar : PubMed/NCBI
38	Lamouille S, Subramanyam D, Blelloch R and Derynck R: Regulation of epithelial-mesenchymal and mesenchymal-epithelial transitions by microRNAs. Curr Opin Cell Biol. 25:200–207. 2013. View Article : Google Scholar : PubMed/NCBI
39	Yu T, Li C, Wang Z, Liu K, Xu C, Yang Q, Tang Y and Wu Y: Non-coding RNAs deregulation in oral squamous cell carcinoma: Advances and challenges. Clin Transl Oncol. 18:427–436. 2016. View Article : Google Scholar : PubMed/NCBI
40	Tu HF, Lin SC and Chang KW: MicroRNA aberrances in head and neck cancer: Pathogenetic and clinical significance. Curr Opin Otolaryngol Head Neck Surg. 21:104–111. 2013. View Article : Google Scholar : PubMed/NCBI
41	Zhang H, Li Y and Lai M: The microRNA network and tumor metastasis. Oncogene. 29:937–948. 2010. View Article : Google Scholar : PubMed/NCBI
42	Bracken CP, Gregory PA, Khew-Goodall Y and Goodall GJ: The role of microRNAs in metastasis and epithelial-mesenchymal transition. Cell Mol Life Sci. 66:1682–1699. 2009. View Article : Google Scholar : PubMed/NCBI
43	Kim KY, Lee GY and Cha IH: Biomarker detection for the diagnosis of lymph node metastasis from oral squamous cell carcinoma. Oral Oncol. 48:311–319. 2012. View Article : Google Scholar : PubMed/NCBI
44	Severino P, Brüggemann H, Andreghetto FM, Camps C, Mde F Klingbeil, de Pereira WO, Soares RM, Moyses R, Wünsch-Filho V, Mathor MB, et al: MicroRNA expression profile in head and neck cancer: HOX-cluster embedded microRNA-196a and microRNA-10b dysregulation implicated in cell proliferation. BMC Cancer. 13:5332013. View Article : Google Scholar : PubMed/NCBI
45	Platais C, Hakami F, Darda L, Lambert DW, Morgan R and Hunter KD: The role of HOX genes in head and neck squamous cell carcinoma. J Oral Pathol Med. 45:239–247. 2016. View Article : Google Scholar : PubMed/NCBI
46	Darda L, Hakami F, Morgan R, Murdoch C, Lambert DW and Hunter KD: The role of HOXB9 and miR-196a in head and neck squamous cell carcinoma. PLoS One. 10:e01222852015. View Article : Google Scholar : PubMed/NCBI
47	Jiang Y, Zhang Y, Li F, Du X and Zhang J: CDX2 inhibits pancreatic adenocarcinoma cell proliferation via promoting tumor suppressor miR-615-5p. Tumour Biol. 37:1041–1049. 2016. View Article : Google Scholar : PubMed/NCBI
48	Pearson CE: Co-opting endogenous microRNAs for therapy. Nat Med. 18:1011–1012. 2012. View Article : Google Scholar : PubMed/NCBI
49	Hoss AG, Kartha VK, Dong X, Latourelle JC, Dumitriu A, Hadzi TC, Macdonald ME, Gusella JF, Akbarian S, Chen JF, et al: MicroRNAs located in the Hox gene clusters are implicated in huntington's disease pathogenesis. PLoS Genet. 10:e10041882014. View Article : Google Scholar : PubMed/NCBI
50	Shan Q, Zheng G, Zhu A, Cao L, Lu J, Wu D, Zhang Z, Fan S, Sun C, Hu B, et al: Epigenetic modification of miR-10a regulates renal damage by targeting CREB1 in type 2 diabetes mellitus. Toxicol Appl Pharmacol. 306:134–143. 2016. View Article : Google Scholar : PubMed/NCBI
51	Bourguignon LY, Wong G and Shiina M: Up-regulation of histone methyltransferase, DOT1L, by matrix hyaluronan promotes MicroRNA-10 expression leading to tumor cell invasion and chemoresistance in cancer stem cells from head and neck squamous cell carcinoma. J Biol Chem. 291:10571–10585. 2016. View Article : Google Scholar : PubMed/NCBI
52	Lu YC, Chang JT, Liao CT, Kang CJ, Huang SF, Chen IH, Huang CC, Huang YC, Chen WH, Tsai CY, et al: OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway. Mol Cancer. 13:2182014. View Article : Google Scholar : PubMed/NCBI
53	Suh YE, Raulf N, Gäken J, Lawler K, Urbano TG, Bullenkamp J, Gobeil S, Huot J, Odell E and Tavassoli M: MicroRNA-196a promotes an oncogenic effect in head and neck cancer cells by suppressing annexin A1 and enhancing radioresistance. Int J Cancer. 137:1021–1034. 2015. View Article : Google Scholar : PubMed/NCBI
54	Saito K, Inagaki K, Kamimoto T, Ito Y, Sugita T, Nakajo S, Hirasawa A, Iwamaru A, Ishikura T, Hanaoka H, et al: MicroRNA-196a is a putative diagnostic biomarker and therapeutic target for laryngeal cancer. PLoS One. 8:e714802013. View Article : Google Scholar : PubMed/NCBI
55	Kozaki K, Imoto I, Mogi S, Omura K and Inazawa J: Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancer. Cancer Res. 68:2094–2105. 2008. View Article : Google Scholar : PubMed/NCBI
56	Lu YC, Chen YJ, Wang HM, Tsai CY, Chen WH, Huang YC, Fan KH, Tsai CN, Huang SF, Kang CJ, et al: Oncogenic function and early detection potential of miRNA-10b in oral cancer as identified by microRNA profiling. Cancer Prev Res (Phila). 5:665–674. 2012. View Article : Google Scholar : PubMed/NCBI
57	Ganci F, Sacconi A, Manciocco V, Sperduti I, Battaglia P, Covello R, Muti P, Strano S, Spriano G, Fontemaggi G, et al: MicroRNA expression as predictor of local recurrence risk in oral squamous cell carcinoma. Head Neck. 38:E189–E197. 2016. View Article : Google Scholar : PubMed/NCBI
58	Sasahira T, Kurihara M, Bhawal UK, Ueda N, Shimomoto T, Yamamoto K, Kirita T and Kuniyasu H: Downregulation of miR-126 induces angiogenesis and lymphangiogenesis by activation of VEGF-A in oral cancer. Br J Cancer. 107:700–706. 2012. View Article : Google Scholar : PubMed/NCBI
59	Zhang X, Yang H, Lee JJ, Kim E, Lippman SM, Khuri FR, Spitz MR, Lotan R, Hong WK and Wu X: MicroRNA-related genetic variations as predictors for risk of second primary tumor and/or recurrence in patients with early-stage head and neck cancer. Carcinogenesis. 31:2118–2123. 2010. View Article : Google Scholar : PubMed/NCBI
60	Gombos K, Horváth R, Szele E, Juhász K, Gocze K, Somlai K, Pajkos G, Ember I and Olasz L: miRNA expression profiles of oral squamous cell carcinomas. Anticancer Res. 33:1511–1517. 2013.PubMed/NCBI
61	Kina S, Nakasone T, Kinjo T, Maruyama T, Kawano T and Arasaki A: Impact of metronomic neoadjuvant chemotherapy on early tongue cancer. Cancer Chemother Pharmacol. 78:833–840. 2016. View Article : Google Scholar : PubMed/NCBI
62	Peisker A, Raschke GF, Guentsch A, Luepke P, Roshanghias K and Schultze-Mosgau S: Evaluation of a post-treatment follow-up program in patients with oral squamous cell carcinoma. Clin Oral Investig. 21:135–141. 2017. View Article : Google Scholar : PubMed/NCBI
63	Yanamoto S, Yamada S, Takahashi H, Kawasaki G, Ikeda H, Shiraishi T, Fujita S, Ikeda T, Asahina I and Umeda M: Predictors of locoregional recurrence in T1-2N0 tongue cancer patients. Pathol Oncol Res. 19:795–803. 2013. View Article : Google Scholar : PubMed/NCBI
64	Johnson N, Franceschi S, Ferlay J, Ramadas K, Schmid S, MacDonald DG, Bouquot JE and Slootweg PJ: Squamous cell carcinoma. In: World Health Organization Classification of TumoursPathology & Genetics Head and Neck Tumours. Barnes C, Eveson JW, Reichart P and Sidransky D: IARC Press; Lyon: pp. 168–175. 2005
65	Yamamoto E, Kohama G, Sunakawa H, Iwai M and Hiratsuka H: Mode of invasion, bleomycin sensitivity and clinical course in squamous cell carcinoma of the oral cavity. Cancer. 51:2175–2180. 1983. View Article : Google Scholar : PubMed/NCBI
66	Huang SH, Hwang D, Lockwood G, Goldstein DP and O'Sullivan B: Predictive value of tumor thickness for cervical lymph-node involvement in squamous cell carcinoma of the oral cavity: A meta-analysis of reported studies. Cancer. 115:1489–1497. 2009. View Article : Google Scholar : PubMed/NCBI
67	Zeka F, Vanderheyden K, De Smet E, Cuvelier CA, Mestdagh P and Vandesompele J: Straightforward and sensitive RT-qPCR based gene expression analysis of FFPE samples. Sci Rep. 6:214182016. View Article : Google Scholar : PubMed/NCBI
68	Nagao Y, Hisaoka M, Matsuyama A, Kanemitsu S, Hamada T, Fukuyama T, Nakano R, Uchiyama A, Kawamoto M, Yamaguchi K, et al: Association of microRNA-21 expression with its targets, PDCD4 and TIMP3, in pancreatic ductal adenocarcinoma. Mod Pathol. 25:112–121. 2012. View Article : Google Scholar : PubMed/NCBI
69	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
70	Fanale D, Amodeo V, Bazan V, Insalaco L, Incorvaia L, Barraco N, Castiglia M, Rizzo S, Santini D, Giordano A, et al: Can the microRNA expression profile help to identify novel targets for zoledronic acid in breast cancer? Oncotarget. 7:29321–29332. 2016. View Article : Google Scholar : PubMed/NCBI
71	McCall MN, McMurray HR, Land H and Almudevar A: On non-detects in qPCR data. Bioinformatics. 30:2310–2316. 2014. View Article : Google Scholar : PubMed/NCBI
72	Lee RC, Feinbaum RL and Ambros V: The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 75:843–854. 1993. View Article : Google Scholar : PubMed/NCBI
73	Scott SE, Grunfeld EA and McGurk M: The idiosyncratic relationship between diagnostic delay and stage of oral squamous cell carcinoma. Oral Oncol. 41:396–403. 2005. View Article : Google Scholar : PubMed/NCBI
74	Mitani S, Tomioka T, Hayashi R, Ugumori T, Hato N and Fujii S: Anatomic invasive depth predicts delayed cervical lymph node metastasis of tongue squamous cell carcinoma. Am J Surg Pathol. 40:934–942. 2016. View Article : Google Scholar : PubMed/NCBI
75	Orabona GD, Bonavolontà P, Maglitto F, Friscia M, Iaconetta G and Califano L: Neck dissection versus ‘watchful-waiting’ in early squamous cell carcinoma of the tongue our experience on 127 cases. Surg Oncol. 25:401–404. 2016. View Article : Google Scholar : PubMed/NCBI
76	D'Cruz AK, Vaish R, Kapre N, Dandekar M, Gupta S, Hawaldar R, Agarwal JP, Pantvaidya G, Chaukar D, Deshmukh A, et al: Elective versus therapeutic neck dissection in node-negative oral cancer. N Engl J Med. 373:521–529. 2015. View Article : Google Scholar : PubMed/NCBI
77	Irani S: miRNAs signature in head and neck squamous cell carcinoma metastasis: A literature review. J Dent (Shiraz). 17:71–83. 2016.PubMed/NCBI
78	Ganly I, Goldstein D, Carlson DL, Patel SG, O'Sullivan B, Lee N, Gullane P and Shah JP: Long-term regional control and survival in patients with ‘low-risk,’ early stage oral tongue cancer managed by partial glossectomy and neck dissection without postoperative radiation: The importance of tumor thickness. Cancer. 119:1168–1176. 2013. View Article : Google Scholar : PubMed/NCBI
79	Hebert C, Norris K, Scheper MA, Nikitakis N and Sauk JJ: High mobility group A2 is a target for miRNA-98 in head and neck squamous cell carcinoma. Mol Cancer. 6:52007. View Article : Google Scholar : PubMed/NCBI
80	Yu T, Wang XY, Gong RG, Li A, Yang S, Cao YT, Wen YM, Wang CM and Yi XZ: The expression profile of microRNAs in a model of 7,12-dimethyl-benz [a]anthrance-induced oral carcinogenesis in Syrian hamster. J Exp Clin Cancer Res. 28:642009. View Article : Google Scholar : PubMed/NCBI
81	Wong TS, Liu XB, Chung-Wai Ho A, Po-Wing Yuen A, Wai-Man Ng R and Ignace Wei W: Identification of pyruvate kinase type M2 as potential oncoprotein in squamous cell carcinoma of tongue through microRNA profiling. Int J Cancer. 123:251–257. 2008. View Article : Google Scholar : PubMed/NCBI
82	Grizzi F, Di Ieva A, Russo C, Frezza EE, Cobos E, Muzzio PC and Chiriva-Internati M: Cancer initiation and progression: An unsimplifiable complexity. Theor Biol Med Model. 3:372006. View Article : Google Scholar : PubMed/NCBI
83	Brito BL, Lourenço SV, Damascena AS, Kowalski LP, Soares FA and Coutinho-Camillo CM: Expression of stem cell-regulating miRNAs in oral cavity and oropharynx squamous cell carcinoma. J Oral Pathol Med. 45:647–654. 2016. View Article : Google Scholar : PubMed/NCBI
84	Manikandan M, Magendhra Deva Rao AK, Rajkumar KS, Rajaraman R and Munirajan AK: Altered levels of miR-21, miR-125b-2*, miR-138, miR-155, miR-184 and miR-205 in oral squamous cell carcinoma and association with clinicopathological characteristics. J Oral Pathol Med. 44:792–800. 2015. View Article : Google Scholar : PubMed/NCBI
85	Tu HF, Chang KW, Cheng HW and Liu CJ: Upregulation of miR-372 and −373 associates with lymph node metastasis and poor prognosis of oral carcinomas. Laryngoscope. 125:E365–E370. 2015. View Article : Google Scholar : PubMed/NCBI
86	Chang WM, Lin YF, Su CY, Peng HY, Chang YC, Lai TC, Wu GH, Hsu YM, Chi LH, Hsiao JR, et al: Dysregulation of RUNX2/activin-A axis upon miR-376c downregulation promotes lymph node metastasis in head and neck squamous cell carcinoma. Cancer Res. 76:7140–7150. 2016. View Article : Google Scholar : PubMed/NCBI
87	Peng SC, Liao CT, Peng CH, Cheng AJ, Chen SJ, Huang CG, Hsieh WP and Yen TC: MicroRNAs MiR-218, MiR-125b, and Let-7g predict prognosis in patients with oral cavity squamous cell carcinoma. PLoS One. 9:e1024032014. View Article : Google Scholar : PubMed/NCBI
88	Chang CC, Yang YJ, Li YJ, Chen ST, Lin BR, Wu TS, Lin SK, Kuo MY and Tan CT: MicroRNA-17/20a functions to inhibit cell migration and can be used a prognostic marker in oral squamous cell carcinoma. Oral Oncol. 49:923–931. 2013. View Article : Google Scholar : PubMed/NCBI
89	Li J, Huang H, Sun L, Yang M, Pan C, Chen W, Wu D, Lin Z, Zeng C, Yao Y, et al: MiR-21 indicates poor prognosis in tongue squamous cell carcinomas as an apoptosis inhibitor. Clin Cancer Res. 15:3998–4008. 2009. View Article : Google Scholar : PubMed/NCBI
90	Cao J, Guo T, Dong Q, Zhang J and Li Y: miR-26b is downregulated in human tongue squamous cell carcinoma and regulates cell proliferation and metastasis through a COX-2-dependent mechanism. Oncol Rep. 33:974–980. 2015. View Article : Google Scholar : PubMed/NCBI
91	Yang CN, Deng YT, Tang JY, Cheng SJ, Chen ST, Li YJ, Wu TS, Yang MH, Lin BR, Kuo MY, et al: MicroRNA-29b regulates migration in oral squamous cell carcinoma and its clinical significance. Oral Oncol. 51:170–177. 2015. View Article : Google Scholar : PubMed/NCBI
92	Siow MY, Ng LP, Vincent-Chong VK, Jamaludin M, Abraham MT, Rahman Abdul ZA, Kallarakkal TG, Yang YH, Cheong SC and Zain RB: Dysregulation of miR-31 and miR-375 expression is associated with clinical outcomes in oral carcinoma. Oral Dis. 20:345–351. 2014. View Article : Google Scholar : PubMed/NCBI
93	Jia LF, Wei SB, Mitchelson K, Gao Y, Zheng YF, Meng Z, Gan YH and Yu GY: miR-34a inhibits migration and invasion of tongue squamous cell carcinoma via targeting MMP9 and MMP14. PLoS One. 9:e1084352014. View Article : Google Scholar : PubMed/NCBI
94	Li G, Ren S, Su Z, Liu C, Deng T, Huang D, Tian Y, Qiu Y and Liu Y: Increased expression of miR-93 is associated with poor prognosis in head and neck squamous cell carcinoma. Tumour Biol. 36:3949–3956. 2015. View Article : Google Scholar : PubMed/NCBI
95	Liu CJ, Shen WG, Peng SY, Cheng HW, Kao SY, Lin SC and Chang KW: miR-134 induces oncogenicity and metastasis in head and neck carcinoma through targeting WWOX gene. Int J Cancer. 134:811–821. 2014. View Article : Google Scholar : PubMed/NCBI
96	Xu Q, Sun Q, Zhang J, Yu J, Chen W and Zhang Z: Downregulation of miR-153 contributes to epithelial-mesenchymal transition and tumor metastasis in human epithelial cancer. Carcinogenesis. 34:539–549. 2013. View Article : Google Scholar : PubMed/NCBI
97	Baba O, Hasegawa S, Nagai H, Uchida F, Yamatoji M, Kanno NI, Yamagata K, Sakai S, Yanagawa T and Bukawa H: MicroRNA-155-5p is associated with oral squamous cell carcinoma metastasis and poor prognosis. J Oral Pathol Med. 45:248–255. 2016. View Article : Google Scholar : PubMed/NCBI
98	Yang CC, Hung PS, Wang PW, Liu CJ, Chu TH, Cheng HW and Lin SC: miR-181 as a putative biomarker for lymph-node metastasis of oral squamous cell carcinoma. J Oral Pathol Med. 40:397–404. 2011. View Article : Google Scholar : PubMed/NCBI
99	Obayashi M, Yoshida M, Tsunematsu T, Ogawa I, Sasahira T, Kuniyasu H, Imoto I, Abiko Y, Xu D, Fukunaga S, et al: microRNA-203 suppresses invasion and epithelial-mesenchymal transition induction via targeting NUAK1 in head and neck cancer. Oncotarget. 7:8223–8239. 2016. View Article : Google Scholar : PubMed/NCBI
100	Chang KW, Liu CJ, Chu TH, Cheng HW, Hung PS, Hu WY and Lin SC: Association between high miR-211 microRNA expression and the poor prognosis of oral carcinoma. J Dent Res. 87:1063–1068. 2008. View Article : Google Scholar : PubMed/NCBI
101	Liu X, Yu J, Jiang L, Wang A, Shi F, Ye H and Zhou X: MicroRNA-222 regulates cell invasion by targeting matrix metalloproteinase 1 (MMP1) and manganese superoxide dismutase 2 (SOD2) in tongue squamous cell carcinoma cell lines. Cancer Genomics Proteomics. 6:131–139. 2009.PubMed/NCBI
102	Sun Q, Zhang J, Cao W, Wang X, Xu Q, Yan M, Wu X and Chen W: Dysregulated miR-363 affects head and neck cancer invasion and metastasis by targeting podoplanin. Int J Biochem Cell Biol. 45:513–520. 2013. View Article : Google Scholar : PubMed/NCBI
103	Huang WC, Chan SH, Jang TH, Chang JW, Ko YC, Yen TC, Chiang SL, Chiang WF, Shieh TY, Liao CT, et al: miRNA-491-5p and GIT1 serve as modulators and biomarkers for oral squamous cell carcinoma invasion and metastasis. Cancer Res. 74:751–764. 2014. View Article : Google Scholar : PubMed/NCBI
104	Liao L, Wang J, Ouyang S, Zhang P, Wang J and Zhang M: Expression and clinical significance of microRNA-1246 in human oral squamous cell carcinoma. Med Sci Monit. 21:776–781. 2015. View Article : Google Scholar : PubMed/NCBI
105	Manikandan M, Magendhra Deva Rao AK, Arunkumar G, Manickavasagam M, Rajkumar KS, Rajaraman R and Munirajan AK: Oral squamous cell carcinoma: microRNA expression profiling and integrative analyses for elucidation of tumourigenesis mechanism. Mol Cancer. 15:282016. View Article : Google Scholar : PubMed/NCBI
106	Bhatlekar S, Fields JZ and Boman BM: HOX genes and their role in the development of human cancers. J Mol Med (Berl). 92:811–823. 2014. View Article : Google Scholar : PubMed/NCBI
107	Marcinkiewicz KM and Gudas LJ: Altered epigenetic regulation of homeobox genes in human oral squamous cell carcinoma cells. Exp Cell Res. 320:128–143. 2014. View Article : Google Scholar : PubMed/NCBI
108	Chen C, Zhang Y, Zhang L, Weakley SM and Yao Q: MicroRNA-196: Critical roles and clinical applications in development and cancer. J Cell Mol Med. 15:14–23. 2011. View Article : Google Scholar : PubMed/NCBI
109	Liu Y, Zhang Y, Wu H, Li Y, Zhang Y, Liu M, Li X and Tang H: miR-10a suppresses colorectal cancer metastasis by modulating the epithelial-to-mesenchymal transition and anoikis. Cell Death Dis. 8:e27392017. View Article : Google Scholar : PubMed/NCBI
110	Chen W, Tang Z, Sun Y, Zhang Y, Wang X, Shen Z, Liu F and Qin X: miRNA expression profile in primary gastric cancers and paired lymph node metastases indicates that miR-10a plays a role in metastasis from primary gastric cancer to lymph nodes. Exp Ther Med. 3:351–356. 2012. View Article : Google Scholar : PubMed/NCBI
111	Harris T, Jimenez L, Kawachi N, Fan JB, Chen J, Belbin T, Ramnauth A, Loudig O, Keller CE, Smith R, et al: Low-level expression of miR-375 correlates with poor outcome and metastasis while altering the invasive properties of head and neck squamous cell carcinomas. Am J Pathol. 180:917–928. 2012. View Article : Google Scholar : PubMed/NCBI
112	Veit JA, Scheckenbach K, Schuler PJ, Laban S, Wiggenhauser PS, Thierauf J, Klussmann JP and Hoffmann TK: MicroRNA expression in differentially metastasizing tumors of the head and neck: Adenoid cystic versus squamous cell carcinoma. Anticancer Res. 35:1271–1277. 2015.PubMed/NCBI
113	Min A, Zhu C, Peng S, Rajthala S, Costea DE and Sapkota D: MicroRNAs as important players and biomarkers in oral carcinogenesis. Biomed Res Int. 2015:1869042015. View Article : Google Scholar : PubMed/NCBI
114	Zahran F, Ghalwash D, Shaker O, Al-Johani K and Scully C: Salivary microRNAs in oral cancer. Oral Dis. 21:739–747. 2015. View Article : Google Scholar : PubMed/NCBI
115	Johnson JJ, Miller DL, Jiang R, et al: Protease-activated Receptor-2 (PAR-2)-mediated Nf-κB activation suppresses inflammation-associated tumor Suppressor MicroRNAs in oral squamous cell carcinoma. J Biol Chem. 291:6936–6945. 2016. View Article : Google Scholar : PubMed/NCBI
116	Mascolo M, Siano M, Ilardi G, Russo D, Merolla F, De Rosa G and Staibano S: Epigenetic disregulation in oral cancer. Int J Mol Sci. 13:2331–2353. 2012. View Article : Google Scholar : PubMed/NCBI
117	Hayashi M, Wu G, Roh JL, et al: Correlation of gene methylation in surgical margin imprints with locoregional recurrence in head and neck squamous cell carcinoma. Cancer. 121:1957–1965. 2015. View Article : Google Scholar : PubMed/NCBI
118	Asli NS and Kessel M: Spatiotemporally restricted regulation of generic motor neuron programs by miR-196-mediated repression of Hoxb8. Dev Biol. 344:857–868. 2010. View Article : Google Scholar : PubMed/NCBI
119	Dasen JS: Long noncoding RNAs in development: solidifying the Lncs to Hox gene regulation. Cell Rep. 5:1–2. 2013. View Article : Google Scholar : PubMed/NCBI
120	Ma L: Role of miR-10b in breast cancer metastasis. Breast Cancer Res. 12:2102010. View Article : Google Scholar : PubMed/NCBI
121	Wu Y, Zhang L, Zhang L, Wang Y, Li H, Ren X, Wei F, Yu W, Liu T, Wang X, et al: Long non-coding RNA HOTAIR promotes tumor cell invasion and metastasis by recruiting EZH2 and repressing E-cadherin in oral squamous cell carcinoma. Int J Oncol. 46:2586–2594. 2015. View Article : Google Scholar : PubMed/NCBI
122	Adami GR, Tang JL and Markiewicz MR: Improving accuracy of RNA-based diagnosis and prognosis of oral cancer by using noninvasive methods. Oral Oncol. 69:62–67. 2017. View Article : Google Scholar : PubMed/NCBI
123	He Q, Chen Z, Cabay RJ, et al: microRNA-21 and microRNA-375 from oral cytology as biomarkers for oral tongue cancer detection. Oral Oncol. 57:15–20. 2016. View Article : Google Scholar : PubMed/NCBI
124	Koch FP, Kämmerer PW, Kämmerer P, Al-Nawas B and Brieger J: Influence of class M1 glutathione S-transferase (GST Mu) polymorphism on GST M1 gene expression level and tumor size in oral squamous cell carcinoma. Oral Oncol. 46:128–133. 2010. View Article : Google Scholar : PubMed/NCBI
125	Serrao NR, Reid SM and Wilson CC: Establishing detection thresholds for environmental DNA using receiver operator characteristic (ROC) curves. Conservation Genet Resour. 1–8. 2017.
126	Mordhorst LB, Sorbe B and Ahlin C: A study of serum biomarkers associated with relapse of cervical cancer. Anticancer Res. 32:4913–4922. 2012.PubMed/NCBI
127	Hosmer DW Jr, Lemeshow S and Sturdivant RX: Applied Logistic Regression. 3rd edition. John Wiley & Sons; Hoboken, NJ: pp. 173–182. 2013
128	Braoudaki M, Lambrou GI, Giannikou K, et al: miR-15a and miR-24-1 as putative prognostic microRNA signatures for pediatric pilocytic astrocytomas and ependymomas. Tumour Biol. 37:9887–9897. 2016. View Article : Google Scholar : PubMed/NCBI
129	Braoudaki M, Lambrou GI, Papadodima SA, Stefanaki K, Prodromou N and Kanavakis E: MicroRNA expression profiles in pediatric dysembryoplastic neuroepithelial tumors. Med Oncol. 33:52016. View Article : Google Scholar : PubMed/NCBI
130	Hunt EA, Broyles D, Head T and Deo SK: MicroRNA detection: Current technology and research strategies. Annu Rev Anal Chem (Palo Alto Calif). 8:217–237. 2015. View Article : Google Scholar : PubMed/NCBI
131	Doleshal M, Magotra AA, Choudhury B, Cannon BD, Labourier E and Szafranska AE: Evaluation and validation of total RNA extraction methods for microRNA expression analyses in formalin-fixed, paraffin-embedded tissues. J Mol Diagn. 10:203–211. 2008. View Article : Google Scholar : PubMed/NCBI
132	Dai L, Wang Y, Chen L, Zheng J, Li J and Wu X: MiR-221, a potential prognostic biomarker for recurrence in papillary thyroid cancer. World J Surg Oncol. 15:112017. View Article : Google Scholar : PubMed/NCBI
133	Li J, Smyth P, Flavin R, Cahill S, Denning K, Aherne S, Guenther SM, O'Leary JJ and Sheils O: Comparison of miRNA expression patterns using total RNA extracted from matched samples of formalin-fixed paraffin-embedded (FFPE) cells and snap frozen cells. BMC Biotechnol. 7:362007. View Article : Google Scholar : PubMed/NCBI
134	Manikandan M, Magendhra Deva Rao AK, et al: Down regulation of miR-34a and miR-143 may indirectly inhibit p53 in oral squamous cell carcinoma: A pilot study. Asian Pac J Cancer Prev. 16:7619–7625. 2015. View Article : Google Scholar : PubMed/NCBI
135	Moratin J, Hartmann S, Brands R, et al: Evaluation of miRNA-expression and clinical tumour parameters in oral squamous cell carcinoma (OSCC). J Craniomaxillofac Surg. 44:876–881. 2016. View Article : Google Scholar : PubMed/NCBI