Distinct pattern of expression of differentiation and growth-related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays
Distinct pattern of expression of differentiation and growth-related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays
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DOI：
10.1038/sj.onc.1203703
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发表时间：
2000-06-29
期刊：
影响因子：
8
通讯作者：
Gutkind, JS
中科院分区：
文献类型：
--
作者：
Leethanakul, C;Patel, V;Gutkind, JS
尽管头颈部鳞状细胞癌（HNSCC）的危险因素已经得到了很好的认识，但对这种恶性肿瘤的分子机制知之甚少。此外，研究肿瘤进展不同阶段的基因表达谱的能力通常受到这些肿瘤病变显著异质性的限制。在这里，我们展示了激光捕获显微解剖（LCM）的成功使用，以获得特定的细胞群。从具有代表性的HNSCC及其匹配的正常组织中提取的5000个细胞足以提取高完整性的RNA，用于合成标记扩增的cDNA探针，然后将其杂交到这些排列有已知人类癌症相关cDNA的膜上。此外，与正常组织相比，我们发现分化标志物（如细胞角蛋白）的表达持续下降，而一些信号转导和细胞周期调节分子，以及生长和血管生成因子和组织降解蛋白酶的表达增加。出乎意料的是，我们还发现大多数HNSCC过表达wnt和notch生长和分化调控系统的成员，从而表明，wnt和notch通路可能参与鳞状细胞癌变。这种实验方法可能有助于识别候选标记物，用于早期发现肿瘤前病变，以及对该疾病进行药物干预的新靶点。
Although risk factors for squamous cell carcinomas of the head and neck (HNSCC) are well recognized, very little is known about the molecular mechanisms responsible for this malignancy. Furthermore, the ability to investigate gene expression profiles at different stages of tumor progression is usually limited by the remarkable heterogeneity of these neoplastic lesions. Here, we show the successful use of laser capture microdissection (LCM) to procure specific cell populations. The 5000 cells from representative sets of HNSCC and their matching normal tissues are sufficient to extract RNA of high integrity for the synthesis of labeled amplified cDNA probes which can then be hybridized to these membranes arrayed with known human cancer-related cDNAs. Furthermore, when compared to normal tissues, we demonstrate a consistent decrease in expression of differentiation markers such as cytokeratins, and an increase in the expression of a number of signal transducing and cell cycle regulatory molecules, as well as growth and angiogenic factors and tissue degrading proteases. Unexpectedly, we also found that most HNSCC overexpress members of the wnt and notch growth and differentiation regulatory system, thus suggesting that the,wnt and notch pathways mag contribute in squamous cell carcinogenesis. This experimental approach may facilitate the identification candidate markers for the early detection of preneoplastic lesions, as well as novel targets for pharmacological intervention in this disease.