Identification and characterization of dysregulated P‑element induced wimpy testis‑interacting RNAs in head and neck squamous cell carcinoma

Saad,Maarouf  A.; Ku,Jonjei; Kuo,Selena  Z.; Li,Pin  Xue; Zheng,Hao; Yu,Michael  Andrew; Wang‑Rodriguez,Jessica; Ongkeko,Weg  M.

doi:10.3892/ol.2019.9913

Identification and characterization of dysregulated P‑element induced wimpy testis‑interacting RNAs in head and neck squamous cell carcinoma

Authors:
- Maarouf A. Saad
- Jonjei Ku
- Selena Z. Kuo
- Pin Xue Li
- Hao Zheng
- Michael Andrew Yu
- Jessica Wang‑Rodriguez
- Weg M. Ongkeko
View Affiliations

Affiliations: School of Medicine, Yale University, New Haven, CT 06510, USA, Division of Otolaryngology‑Head and Neck Surgery, Department of Surgery, University of California, San Diego, La Jolla, CA 92093, USA, School of Medicine, University of California, San Francisco, CA 94143, USA, Veterans Administration Medical Center and Department of Pathology, University of California, San Diego, La Jolla, CA 92161, USA
Published online on: January 9, 2019 https://doi.org/10.3892/ol.2019.9913
Pages: 2615-2622
Copyright: © Saad et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

It is clear that alcohol consumption is a major risk factor in the pathogenesis of head and neck squamous cell carcinoma (HNSCC); however, the molecular mechanism underlying the pathogenesis of alcohol‑associated HNSCC remains poorly understood. The aim of the present study was to identify and characterize P‑element‑induced wimpy testis (PIWI)‑interacting RNAs (piRNAs) and PIWI proteins dysregulated in alcohol‑associated HNSCC to elucidate their function in the development of this cancer. Using next generation RNA‑sequencing (RNA‑seq) data obtained from 40 HNSCC patients, the piRNA and PIWI protein expression of HNSCC samples was compared between alcohol drinkers and non‑drinkers. A separate piRNA expression RNA‑seq analysis of 18 non‑smoker HNSCC patients was also conducted. To verify piRNA expression, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed on the most differentially expressed alcohol‑associated piRNAs in ethanol and acetaldehyde‑treated normal oral keratinocytes. The correlation between piRNA expression and patient survival was analyzed using Kaplan‑Meier estimators and multivariate Cox proportional hazard models. A comparison between alcohol drinking and non‑drinking HNSCC patients demonstrated that a panel of 3,223 piRNA transcripts were consistently detected and differentially expressed. RNA‑seq analysis and in vitro RT‑qPCR verification revealed that 4 of these piRNAs, piR‑35373, piR‑266308, piR‑58510 and piR‑38034, were significantly dysregulated between drinking and non‑drinking cohorts. Of these four piRNAs, low expression of piR‑58510 and piR‑35373 significantly correlated with improved patient survival. Furthermore, human PIWI‑like protein 4 was consistently upregulated in ethanol and acetaldehyde‑treated normal oral keratinocytes. These results demonstrate that alcohol consumption may cause dysregulation of piRNA expression in HNSCC and in vitro verifications identified 4 piRNAs that may be involved in the pathogenesis of alcohol‑associated HNSCC.

View Figures

View References

1	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar : PubMed/NCBI
2	Nemunaitis J and O'Brien J: Head and neck cancer: Gene therapy approaches. Part 1: Adenoviral vectors. Expert Opin Biol Ther. 2:177–185. 2002. View Article : Google Scholar : PubMed/NCBI
3	Silveira NJ, Varuzza L, Machado-Lima A, Lauretto MS, Pinheiro DG, Rodrigues RV, Severino P, Nobrega FG; Head and Neck Genome Project GENCAPO, ; Silva WA Jr, et al: Searching for molecular markers in head and neck squamous cell carcinomas (HNSCC) by statistical and bioinformatic analysis of larynx-derived SAGE libraries. BMC Med Genomics. 1:562008. View Article : Google Scholar : PubMed/NCBI
4	Argiris A, Karamouzis MV, Raben D and Ferris RL: Head and neck cancer. Lancet. 371:1695–1709. 2008. View Article : Google Scholar : PubMed/NCBI
5	Hashibe M, Brennan P, Benhamou S, Castellsague X, Chen C, Curado MP, Dal Maso L, Daudt AW, Fabianova E, Fernandez L, et al: Alcohol drinking in never users of tobacco, cigarette smoking in never drinkers, and the risk of head and neck cancer: Pooled analysis in the international head and neck cancer epidemiology consortium. J Natl Cancer Inst. 99:777–789. 2007. View Article : Google Scholar : PubMed/NCBI
6	Bobo JK and Husten C: Sociocultural influences on smoking and drinking. Alcohol Res Health. 24:225–232. 2000.PubMed/NCBI
7	Sturgis EM, Wei Q and Spitz MR: Descriptive epidemiology and risk factors for head and neck cancer. Semin Oncol. 31:726–733. 2004. View Article : Google Scholar : PubMed/NCBI
8	Jithesh PV, Risk JM, Schache AG, Dhanda J, Lane B, Liloglou T and Shaw RJ: The epigenetic landscape of oral squamous cell carcinoma. Br J Cancer. 108:370–379. 2013. View Article : Google Scholar : PubMed/NCBI
9	Worsham MJ, Chen KM, Ghanem T, Stephen JK and Divine G: Epigenetic modulation of signal transduction pathways in HPV-associated HNSCC. Otolaryngol Head Neck Surg. 149:409–416. 2013. View Article : Google Scholar : PubMed/NCBI
10	Huang T, Alvarez A, Hu B and Cheng SY: Noncoding RNAs in cancer and cancer stem cells. Chin J Cancer. 32:582–593. 2013. View Article : Google Scholar : PubMed/NCBI
11	Girard A, Sachidanandam R, Hannon GJ and Carmell MA: A germline-specific class of small RNAs binds mammalian Piwi proteins. Nature. 442:199–202. 2006.PubMed/NCBI
12	Esteller M: Non-coding RNAs in human disease. Nat Rev Genet. 12:861–874. 2011. View Article : Google Scholar : PubMed/NCBI
13	Galasso M, Sana ME and Volinia S: Non-coding RNAs: A key to future personalized molecular therapy? Genome Med. 2(12)2010.PubMed/NCBI
14	Mei Y, Clark D and Mao L: Novel dimensions of piRNAs in cancer. Cancer Lett. 336:46–52. 2013. View Article : Google Scholar : PubMed/NCBI
15	Cheng J, Deng H, Xiao B, Zhou H, Zhou F, Shen Z and Guo J: piR-823, a novel non-coding small RNA, demonstrates in vitro and in vivo tumor suppressive activity in human gastric cancer cells. Cancer Lett. 315:12–17. 2012. View Article : Google Scholar : PubMed/NCBI
16	Cheng J, Guo JM, Xiao BX, Miao Y, Jiang Z, Zhou H and Li QN: piRNA, the new non-coding RNA, is aberrantly expressed in human cancer cells. Clin Chim Acta. 412:1621–1625. 2011. View Article : Google Scholar : PubMed/NCBI
17	Huang G, Hu H, Xue X, Shen S, Gao E, Guo G, Shen X and Zhang X: Altered expression of piRNAs and their relation with clinicopathologic features of breast cancer. Clin Transl Oncol. 15:563–568. 2013. View Article : Google Scholar : PubMed/NCBI
18	Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, Pimentel H, Salzberg SL, Rinn JL and Pachter L: Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc. 7:562–578. 2012. View Article : Google Scholar : PubMed/NCBI
19	Homann N, Tillonen J, Meurman JH, Rintamäki H, Lindqvist C, Rautio M, Jousimies-Somer H and Salaspuro M: Increased salivary acetaldehyde levels in heavy drinkers and smokers: A microbiological approach to oral cavity cancer. Carcinogenesis. 21:663–668. 2000. View Article : Google Scholar : PubMed/NCBI
20	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
21	Kiang A, Yu MA and Ongkeko WM: Progress and pitfalls in the identification of cancer stem cell-targeting therapies in head and neck squamous cell carcinoma. Curr Med Chem. 19:6056–6064. 2012. View Article : Google Scholar : PubMed/NCBI
22	Matta A and Ralhan R: Overview of current and future biologically based targeted therapies in head and neck squamous cell carcinoma. Head Neck Oncol. 1:62009. View Article : Google Scholar : PubMed/NCBI
23	Kleiber ML, Diehl EJ, Laufer BI, Mantha K, Chokroborty-Hoque A, Alberry B and Singh SM: Long-term genomic and epigenomic dysregulation as a consequence of prenatal alcohol exposure: A model for fetal alcohol spectrum disorders. Front Genet. 5:1612014. View Article : Google Scholar : PubMed/NCBI
24	Maccani MA and Knopik VS: Cigarette smoke exposure-associated alterations to non-coding RNA. Front Genet. 3:532012. View Article : Google Scholar : PubMed/NCBI
25	Aalto AP and Pasquinelli AE: Small non-coding RNAs mount a silent revolution in gene expression. Curr Opin Cell Biol. 24:333–340. 2012. View Article : Google Scholar : PubMed/NCBI
26	Siomi MC, Sato K, Pezic D and Aravin AA: PIWI-interacting small RNAs: The vanguard of genome defence. Nat Rev Mol Cell Biol. 12:246–258. 2011. View Article : Google Scholar : PubMed/NCBI
27	Kim SK, Jung WH and Koo JS: Differential expression of enzymes associated with serine/glycine metabolism in different breast cancer subtypes. PLoS One. 9:e1010042014. View Article : Google Scholar : PubMed/NCBI
28	Kim YH, Jung WH and Koo JS: Expression of metabolism-related proteins in invasive lobular carcinoma: Comparison to invasive ductal carcinoma. Tumour Biol. 35:10381–10393. 2014. View Article : Google Scholar : PubMed/NCBI