口腔癌中的唾液微小RNA
Salivary microRNAs in oral cancer.
作者信息
Zahran F, Ghalwash D, Shaker O, Al-Johani K, Scully C
机构信息
Division of Oral Medicine, Oral Diagnostic Sciences Department, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia.
Department of Oral Medicine and Periodontology, Faculty of Oral and Dental Medicine, Cairo University, Cairo, Egypt.
出版信息
Oral Dis. 2015 Sep;21(6):739-47. doi: 10.1111/odi.12340. Epub 2015 Apr 22.
OBJECTIVE
This study investigated the use of three salivary microRNAs (miRNA-21, miRNA-184, and miRNA-145) as possible markers for malignant transformation in oral mucosal lesions.
MATERIALS AND METHODS
Salivary whole unstimulated samples were collected from a study group of 100 subjects, consisting of 20 clinically healthy controls, 40 patients with oral potentially malignant disorders (PMDs) [20 with dysplastic lesions and 20 without dysplasia], 20 with biopsy-confirmed oral squamous cell carcinoma (OSCC), and 20 with recurrent aphthous stomatitis (RAS) as disease controls. Total RNA was isolated and purified from saliva samples using the microRNA Isolation Kit (Qiagen, UL). miRNA expression analysis was performed using qRT-PCR (Applied Biosystems).
RESULTS
There was a highly significant increase in salivary miRNA-21 and miRNA-184 in OSCC and PMD (with and without dysplasia) when compared to healthy and disease controls (P < 0.001). Conversely, miRNA-145 levels showed a highly significant decrease in OSCC and PMD overall (P < 0.001). RAS cases showed no significant difference from normal controls in any measured miRNA (P > 0.05). The only microRNA to discriminate between OSCC and PMD with dysplasia was miRNA-184. When receiver operating characteristic curves were designed for the three miRNAs, cutoff points delineating the occurrence of malignant change were a fourfold increase in miRNA-21 with specificity 65% and sensitivity 65%, a 0.6 decrease in miRNA-145, with specificity 70% and sensitivity 60%, and a threefold increase of miRNA-184, with specificity 75% and sensitivity 80%. Calculating the area under the curve revealed that miRNA-184 was the only one among the studied miRNAs that provided good diagnostic value.
CONCLUSION
Salivary determination of the miRNAs tested might furnish a noninvasive, rapid adjunctive aid for revealing malignant transformation in oral mucosal lesions, particularly miRNA-184.
目的
本研究调查了三种唾液微小RNA(miRNA-21、miRNA-184和miRNA-145)作为口腔黏膜病变恶性转化潜在标志物的应用情况。
材料与方法
从100名受试者的研究组中收集未刺激的全唾液样本,该研究组包括20名临床健康对照者、40名口腔潜在恶性疾病(PMD)患者[20名有发育异常病变者和20名无发育异常者]、20名经活检确诊的口腔鳞状细胞癌(OSCC)患者以及20名复发性阿弗他口炎(RAS)患者作为疾病对照。使用微小RNA分离试剂盒(Qiagen,UL)从唾液样本中分离并纯化总RNA。使用qRT-PCR(Applied Biosystems)进行miRNA表达分析。
结果
与健康对照者和疾病对照者相比,OSCC以及PMD(有和无发育异常)患者的唾液miRNA-21和miRNA-184显著升高(P < 0.001)。相反,总体上OSCC和PMD患者的miRNA-145水平显著降低(P < 0.001)。RAS病例在任何检测的miRNA中与正常对照者均无显著差异(P > 0.05)。唯一能区分OSCC和有发育异常的PMD的微小RNA是miRNA-184。当为这三种miRNA绘制受试者工作特征曲线时,界定恶性变化发生的截断点为:miRNA-21增加四倍,特异性为65%,敏感性为65%;miRNA-145降低0.6,特异性为70%,敏感性为60%;miRNA-184增加三倍,特异性为75%,敏感性为80%。计算曲线下面积显示,miRNA-184是所研究的miRNA中唯一具有良好诊断价值的。
结论
检测所测试的miRNA的唾液水平可能为揭示口腔黏膜病变的恶性转化提供一种非侵入性、快速的辅助手段,尤其是miRNA-184。