Department of Stomatology, First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Pathol Oncol Res. 2011 Sep;17(3):765-9. doi: 10.1007/s12253-011-9384-9. Epub 2011 May 9.
Recent studies have determined that gene expression profiling using microarray technology can be used to identify tumor-related molecules. The objective of this study was to screen the differentially expressed genes between pleomorphic adenoma (PA) and the normal tissue adjacent to PA using cDNA microarrays and to further validate the differentially expressed genes by real-time PCR. In this study, we selected five pairs of PA and the surrounding normal salivary gland tissues. The total RNA was isolated from tumor and normal tissues and purified to mRNA. The mRNA was reverse-transcribed to cDNA with the incorporation of fluorescent-labeled dUTP to prepare the hybridization probes. The mixed probes were hybridized to Whole Human Gene Expression Microarrays by Agilent. Tumor-related genes were screened by analyzing the fluorescence intensity. As a result, a total of 447 genes were found to be differentially expressed between PA and normal tissue adjacent to PA. Among them, 185 genes were up-regulated and 262 genes were down-regulated in PA. By constructing a network from the differentially expressed genes, some genes, such as Gli2 and CTNNB1, were identified as being at the core of the network. In addition, differential gene expression was validated for 2 up-regulated genes, Gli2 and LOX, using real-time PCR and the results were consistent with those of the cDNA microarray analysis thus verifying the credibility of the microarray data. Therefore, our microarray data may provide clues for finding novel genes involved in the development of PA, and shed light on finding new targets for diagnosis and therapy of PA. Further characterization of these differentially expressed genes will be useful in understanding the genetic basis for PA.
最近的研究已经确定，使用微阵列技术进行基因表达谱分析可以用于鉴定与肿瘤相关的分子。本研究的目的是使用 cDNA 微阵列筛选多形性腺瘤 (PA) 和 PA 周围正常组织之间差异表达的基因，并通过实时 PCR 进一步验证差异表达的基因。在这项研究中，我们选择了五对 PA 和周围正常涎腺组织。从肿瘤和正常组织中分离总 RNA 并纯化至 mRNA。将 mRNA 逆转录为 cDNA，并用荧光标记的 dUTP 掺入制备杂交探针。将混合探针与 Agilent 的全人类基因表达微阵列杂交。通过分析荧光强度筛选肿瘤相关基因。结果，在 PA 和 PA 周围正常组织之间发现了总共 447 个差异表达的基因。其中，PA 中 185 个基因上调，262 个基因下调。通过构建差异表达基因的网络，鉴定出一些基因，如 Gli2 和 CTNNB1，处于网络的核心位置。此外，使用实时 PCR 对 2 个上调基因Gli2 和 LOX 进行了差异基因表达验证，结果与 cDNA 微阵列分析一致，从而验证了微阵列数据的可信度。因此，我们的微阵列数据可能为寻找参与 PA 发生的新基因提供线索，并为寻找 PA 的诊断和治疗新靶点提供线索。对这些差异表达基因的进一步表征将有助于理解 PA 的遗传基础。
