Wong Thian-Sze, Liu Xiao-Bing, Wong Birgitta Yee-Hang, Ng Raymond Wai-Man, Yuen Anthony Po-Wing, Wei William Ignace
Department of Surgery, Faculty of Medicine, The University of Hong Kong, Hong Kong SAR China.
Clin Cancer Res. 2008 May 1;14(9):2588-92. doi: 10.1158/1078-0432.CCR-07-0666.
The aim of this study was to evaluate the microRNA expression patterns in squamous cell carcinoma (SCC) of the tongue.
Expression levels of 156 human mature microRNAs were examined using real-time quantitative PCR (Taq Man MicroRNA Assays; Human Panel) on laser microdissected cells of 4 tongue carcinomas and paired normal tissues. Expression of mature miR-184 was further validated in 20 paired tongue SCC and the normal tissues. Potential oncogenic functions of miR-184 were evaluated in tongue SCC cell lines (Cal27, HN21B, and HN96) with miR-184 inhibitor. Plasma miR-184 levels were evaluated using real-time quantitative PCR.
Using 3-fold expression difference as a cutoff level, we identified 24 up-regulated mature miRNAs including miR-184, miR-34c, miR-137, miR-372, miR-124a, miR-21, miR-124b, miR-31, miR-128a, miR-34b, miR-154, miR-197, miR-132, miR-147, miR-325, miR-181c, miR-198, miR-155, miR-30a-3p, miR-338, miR-17-5p, miR-104, miR-134, and miR-213; and 13 down-regulated mature miRNAs including miR-133a, miR-99a, miR-194, miR-133b, miR-219, miR-100, miR-125b, miR-26b, miR-138, miR-149, miR-195, miR-107, and miR-139. Overexpression of miR-184 was further validated in 20 paired tongue SCC and normal tissues (P = 0.002). Inhibition of miR-184 in tongue SCC cell lines could reduce cell proliferation rate. Down-regulation of c-Myc was observed in two cell lines in response to miR-184 inhibitor. Suppressing miR-184 could induce apoptosis in all three cell lines. Plasma miR-184 levels were significantly higher in tongue SCC patients in comparison with normal individuals, and the levels were significantly reduced after surgical removal of the primary tumors.
Overexpression of miR-184 might play an oncogenic role in the antiapoptotic and proliferative processes of tongue SCC. In addition, plasma miR-184 levels were associated with the presence of primary tumor. Further studies on the aberrantly expressed miRNAs in tongue SCC as well as using plasma miRNAs as novel tumor markers are warranted.
本研究旨在评估舌鳞状细胞癌（SCC）中的微小RNA表达模式。
使用实时定量PCR（Taq Man微小RNA分析；人类芯片）检测4例舌癌及配对正常组织的激光显微切割细胞中156种人类成熟微小RNA的表达水平。在20对配对的舌SCC和正常组织中进一步验证成熟miR-184的表达。使用miR-184抑制剂在舌SCC细胞系（Cal27、HN21B和HN96）中评估miR-184的潜在致癌功能。使用实时定量PCR评估血浆miR-184水平。
以3倍表达差异作为截断水平，我们鉴定出24种上调的成熟微小RNA，包括miR-184、miR-34c、miR-137、miR-372、miR-124a、miR-21、miR-124b、miR-31、miR-128a、miR-34b、miR-154、miR-197、miR-132、miR-147、miR-325、miR-181c、miR-198、miR-155、miR-30a-3p、miR-338、miR-17-5p、miR-104、miR-134和miR-213；以及13种下调的成熟微小RNA，包括miR-133a、miR-99a、miR-194、miR-133b、miR-219、miR-100、miR-125b、miR-26b、miR-138、miR-149、miR-195、miR-107和miR-139。在20对配对的舌SCC和正常组织中进一步验证了miR-184的过表达（P = 0.002）。抑制舌SCC细胞系中的miR-184可降低细胞增殖率。在两种细胞系中观察到c-Myc因miR-184抑制剂而下调。抑制miR-184可诱导所有三种细胞系凋亡。与正常个体相比，舌SCC患者的血浆miR-184水平显著更高，并且在手术切除原发性肿瘤后水平显著降低。
miR-184的过表达可能在舌SCC的抗凋亡和增殖过程中发挥致癌作用。此外，血浆miR-184水平与原发性肿瘤的存在相关。有必要对舌SCC中异常表达的微小RNA以及将血浆微小RNA用作新型肿瘤标志物进行进一步研究。
