RNA-seq analysis of sporadic and Basal Cell Nevus Syndrome-associated odontogenic keratocysts (OKCs)

The aim of this study was to decode the gene expression program characterizing odontogenic keratocyst (OKC) development, by performing a comprehensive transcriptomics analysis applied on whole OKC tissue derived from sporadic and Basal Cell Nevus Syndrome (BCNS)-associated OKCs, and control samples of pericoronal tissues of impacted teeth (dental follicles, DFs) derived from healthy individuals. Formalin-fixed and paraffin-embedded tissue samples from 6 cases of sporadic OKCs, 6 cases of BSCN-associated OKCs and 6 cases of pericoronal tissues of impacted teeth (dental follicles, DFs) were included in RNA-seq analysis. 150-bp paired-end multiplexed sequencing was implemented using an Illumina Hiseq4000 sequencer (Illumina Inc., San Diego, CA). DFs were selected as the control group, as they represent a source of dental lamina rests, proposed to be a potential tissue of origin of OKC. Transcriptomics analysis of the whole OKC tissue was perfomed, composed of the lining epithelium and the fibrous capsule, as the former harbors the pathognomonic features of OKC and the latter is thought to effect its epithelial structure and biological behavior. Validation of RNA-seq results for selected gene markers via immunohistochemistry was performed in all cases.

本研究旨在解析牙源性角化囊肿（odontogenic keratocyst, OKC）发生的特征性基因表达程序，通过对散发性及基底细胞痣综合征（Basal Cell Nevus Syndrome, BCNS）相关OKC的全组织样本，以及健康个体阻生牙冠周组织（牙囊，dental follicles, DFs）的对照样本开展全面转录组学分析。本研究纳入6例散发性OKC、6例BCNS相关OKC及6例阻生牙冠周组织（牙囊，DFs）的福尔马林固定石蜡包埋组织样本，用于RNA测序（RNA-seq）分析。采用Illumina Hiseq4000测序仪（Illumina公司，加利福尼亚州圣地亚哥）完成150bp双端多重测序。选取牙囊作为对照样本，因其代表牙板剩余的来源组织，而牙板剩余被认为是OKC潜在的组织起源。本研究对包含衬里上皮及纤维囊壁的全OKC组织开展转录组学分析——前者具有OKC的特征性病理表现，后者被认为可影响其上皮结构与生物学行为。针对筛选出的基因标志物，通过免疫组织化学方法对所有病例的RNA测序结果进行了验证。