School of Optometry and Ophthalmology and Eye Hospital, Wenzhou Medical College, Wenzhou, China.
PLoS One. 2011 Jan 28;6(1):e16516. doi: 10.1371/journal.pone.0016516.
Uveal melanoma is the most common primary intraocular tumor in adults in western countries. It is associated with very severe visual morbidity and may lead to distant metastases even after successful treatment of the primary tumor. In order to gain better insight into molecular mechanisms related to tumorigenesis and metastasis of uveal melanoma, we used next-generation sequencing technology (SOLiD, Life Technologies) to acquire global transcriptome alteration between posterior uveal melanoma cells and normal uveal melanocyte.
From mRNAs of the cultured uveal melanoma cells and normal uveal melanocytes, we annotated more than 3.7 × 10(7) and 2.7 × 10(7) sequencing tags based on human Ensembl databases, respectively. For detailed analysis, we chose 5155 well-annotated genes mainly involved in the MAPK signaling pathway, cell cycle, cell adhesion junction, apoptosis, and P53 signaling pathways as well as melanogenesis. In an effort to confirm the authenticity of our sequencing results, we validated twenty-one identically differentially expressed genes by using quantitative real time PCR from cultured cell lines of other posterior uveal melanoma cells and normal uveal melanocytes.
We have identified a large number of potentially interesting genes for biological investigation of uveal melanoma. The expression profiling also provides useful resources for other functional genomic and transcriptome studies. These 21 potential genes could discriminate between uveal melanoma cells and normal uveal melanocyte, which may be indicative of tumorigenesis process. Our results further suggest that high-throughput sequencing technology provides a powerful tool to study mechanisms of tumogenesis in the molecular level.
葡萄膜黑色素瘤是西方国家成年人中最常见的原发性眼内肿瘤。它与严重的视力损害有关，即使原发肿瘤得到成功治疗，也可能导致远处转移。为了更好地了解与葡萄膜黑色素瘤发生和转移相关的分子机制，我们使用下一代测序技术（SOLiD，Life Technologies）在培养的后葡萄膜黑色素瘤细胞和正常葡萄膜黑素细胞之间获取了全转录组改变。
从培养的葡萄膜黑色素瘤细胞和正常葡萄膜黑素细胞的 mRNAs 中，我们分别基于人类 Ensembl 数据库注释了超过 3.7×10(7)和 2.7×10(7)个测序标签。为了进行详细分析，我们选择了 5155 个注释良好的基因，这些基因主要涉及 MAPK 信号通路、细胞周期、细胞黏附连接、细胞凋亡和 P53 信号通路以及黑色素生成。为了证实我们测序结果的真实性，我们使用定量实时 PCR 从其他后葡萄膜黑色素瘤细胞和正常葡萄膜黑素细胞的培养细胞系中验证了二十一个同样差异表达的基因。
我们已经鉴定出了大量可能对葡萄膜黑色素瘤生物学研究感兴趣的基因。表达谱分析也为其他功能基因组和转录组研究提供了有用的资源。这 21 个潜在基因可区分葡萄膜黑色素瘤细胞和正常葡萄膜黑素细胞，这可能表明了肿瘤发生过程。我们的结果进一步表明，高通量测序技术为在分子水平上研究肿瘤发生机制提供了一种强大的工具。
