目的：应用高通量基因芯片技术进行转移性喉鳞状细胞癌（laryngeal squamous cell carcinoma，LSCC）中差异表达长链非编码RNA（long noncoding RNA，lncRNA）的筛选及鉴定，寻找有价值的LSCC 相关候选基因和分子标志物。方法：4 例LSCC组织标本均来自河北医科大学耳鼻咽喉头颈外科生物样本库，病理证实为淋巴结转移的LSCC组织及癌旁黏膜组织。提取标本组织总RNA后，应用SBC-lncRNA 基因芯片检测差异表达的lncRNA 及mRNA，采用差异倍数以及Student's t 法对差异表达基因进行筛选，将差异倍数(linear)≤ 0.5 或者≥2.0 和P< 0.05 判断为差异表达基因。选取特定的差异表达lncRNA，应用qRTPCR技术验证芯片结果的可靠性。结果：通过对转移性LSCC组织和癌旁组织的lncRNA表达谱分析显示，转移性LSCC组织与癌旁组织的lncRNA表达谱存在明显差异,差异表达的lncRNA 3 073 个，其中癌组织中表达上调1 967 个、下调1 106 个；差异表达的mRNA 2 809 个，其中癌组织中表达上调1 791 个、下调1 018 个。差异表达的lncRNA主要参与细胞增殖、凋亡及免疫应答等生物学过程，涉及细胞因子及其受体的相互作用、趋化因子信号通路、细胞周期调节、P53 信号通路等。选取其中10 条具有显著差异表达的lncRNA，在25 例LSCC标本组织中进行qRT-PCR验证，发现表达趋势与芯片检测结果一致。结论：转移性LSCC组织及癌旁组织的lncRNA表达谱存在明显差异，深入分析这些差异表达lncRNA的分子作用机制对阐明转移性LSCC的侵袭转移机制有重要的意义，可为LSCC特异性肿瘤标志物的筛选及有效的靶向治疗提供实验依据。

Objective: To search valuable candidate molecular markers for LSCC by screening and identifying differentially expressed long non-coding RNAs (lncRNAs) in metastatic laryngeal squamous cell carcinoma (LSCC) with high throughput gene microarray technique.Methods: Four pairs of LSCC tissues and corresponding para-cancer tissues that pathologically confirmed with lymph node metastasis were collected from Bio-sample lab of Otorhinolaryngology Department, the Second Hospital of Hebei Medical University. After total RNA extraction, the SBC-lncRNA (human 4x180k) chip assay was then applied to detect the differentially expressed lncRNAs and mRNAs, and Fold-change and Student's t-test methods were used to identify differentially expressed genes; the Fold Change (linear)≤0.5 or ≥2.0, P<0.05 was used to identify the differentially expressed genes. The reliability of the chip results was confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results: The gene expression profiles of metastatic LSCC tissues and corresponding para-cancer tissues were significantly different. There were 3 073 differentially expressed lncRNAs, including 1 967 up-regulated and 1 106 down-regulated lncRNAs in cancer tissues. There were 2 809 differentially expressed mRNAs, including 1 791 up- regulated and 1 018 down-regulated mRNAs in cancer tissues. The differentially expressed lncRNAs were mainly in-volved in cell proliferation and apoptosis, immune response biological process, and were associated with cytokine and cytokine receptor interaction, chemokine signaling pathway, cell cycle regulation, P53 signaling pathway, etc. In addition, 10 significantly differentially expressed lncRNAs were chosen and validated by qRT-PCR in 25 cases of LSCC tissues, and the results were in agree with microarray detection. Conclusions: There were obvious differences in lncRNAs expression between metastasis LSCC and corresponding paracancer tissues; in-depth analysis of these differences may has important significance on clarifying the mechanisms of invasion and metastasis of LSCC, which can provide the theoretical basis for biomarker screening and effective targeted therapy for LSCC.

河北省自然科学基金重点资助项目（No.H2017206391）; 河北省政府资助临床医学优秀人才培养和基础课题项目（No.H2017-46）