Saad Maarouf A, Ku Jonjei, Kuo Selena Z, Li Pin Xue, Zheng Hao, Yu Michael Andrew, Wang-Rodriguez Jessica, Ongkeko Weg M
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.
Oncol Lett. 2019 Mar;17(3):2615-2622. doi: 10.3892/ol.2019.9913. Epub 2019 Jan 9.
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 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 verifications identified 4 piRNAs that may be involved in the pathogenesis of alcohol-associated HNSCC.
显然，饮酒是头颈部鳞状细胞癌（HNSCC）发病机制中的一个主要危险因素；然而，酒精相关HNSCC发病机制的分子机制仍知之甚少。本研究的目的是鉴定和表征在酒精相关HNSCC中失调的P元件诱导的微弱睾丸（PIWI）相互作用RNA（piRNA）和PIWI蛋白，以阐明它们在这种癌症发展中的作用。利用从40例HNSCC患者获得的下一代RNA测序（RNA-seq）数据，比较了饮酒者和非饮酒者之间HNSCC样本的piRNA和PIWI蛋白表达。还对18例非吸烟HNSCC患者进行了单独的piRNA表达RNA-seq分析。为了验证piRNA表达，对乙醇和乙醛处理的正常口腔角质形成细胞中差异表达最明显的酒精相关piRNA进行了逆转录定量聚合酶链反应（RT-qPCR）。使用Kaplan-Meier估计器和多变量Cox比例风险模型分析了piRNA表达与患者生存之间的相关性。饮酒和不饮酒的HNSCC患者之间的比较表明，一组3223个piRNA转录本被一致检测到并差异表达。RNA-seq分析和RT-qPCR验证显示，其中4个piRNA，即piR-35373、piR-266308、piR-58510和piR-38034，在饮酒和不饮酒队列之间存在显著失调。在这四个piRNA中，piR-58510和piR-35373的低表达与患者生存率的提高显著相关。此外，人PIWI样蛋白4在乙醇和乙醛处理的正常口腔角质形成细胞中持续上调。这些结果表明，饮酒可能导致HNSCC中piRNA表达失调，验证确定了4个可能参与酒精相关HNSCC发病机制的piRNA。
