应用RNA-Seq技术筛选唾液腺腺样囊性癌嗜神经侵袭相关差异表达基因

doi:10.19438/j.cjoms.2018.02.004

中国口腔颌面外科杂志 ›› 2018, Vol. 16 ›› Issue (2): 114-119.doi: 10.19438/j.cjoms.2018.02.004

应用RNA-Seq技术筛选唾液腺腺样囊性癌嗜神经侵袭相关差异表达基因

李欢, 杨新杰, 王维戚, 雷德林

第四军医大学口腔医院军事口腔医学国家重点实验室,陕西西安 710032;

收稿日期:2017-06-25 修回日期:2017-10-26 出版日期:2018-03-20 发布日期:2018-04-08
通讯作者: 雷德林,E-mail：leidelin@fmmu.edu.cn
作者简介:李欢（1992- ）,男,在读硕士研究生,E-mail：784462885@qq.com
基金资助:
国家自然科学基金（81372901,81302352）

Primary study of the perineural invasion related genes of salivary adenoid cystic carcinoma by RNA-Seq

LI Huan, YANG Xin-jie, WANG Wei-qi, LEI De-lin

State Key Laboratory of Military Stomatology, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University. Xi'an 710032, Shaanxi Province, China;

Received:2017-06-25 Revised:2017-10-26 Online:2018-03-20 Published:2018-04-08

摘要/Abstract

摘要： 目的: 应用RNA-Seq技术分析与施万细胞共培养前、后的唾液腺腺样囊性癌细胞（SACC）的基因表达水平变化,明确嗜神经侵袭（perineural invasion,PNI）相关基因。 方法: 采用腺样囊性癌与SD大鼠施万细胞共培养模型,分析共培养前、后SACC细胞基因表达变化,对差异表达基因进行聚类分析、GO功能富集分析、KEGG通路富集分析,并采用qRT-PCR对其中6个关键差异表达基因进行验证。采用edgeR软件（3.12.1）进行表达差异显著性分析,将P≤0.05、差异表达倍数的绝对值大于1作为差异显著性标准。 结果: 在腺样囊性癌单独培养组与共培养组之间发现395个差异表达基因（P≤0.05,|logFC|≥1.0）,其中135个基因上调（34.2%）,260个基因下调（65.8%）。GO功能富集分析结果表明,这些PNI相关差异表达基因参与了血管再生、细胞外基质的组成、细胞增殖、凋亡和上皮形态发生;KEGG通路富集分析结果表明,这些基因参与组氨酸代谢、肿瘤坏死因子、趋化因子等细胞因子受体相互作用等重要生物学通路。利用qRT-PCR技术对其中6个关键基因进行验证,验证结果与RNA-Seq趋势一致。 结论: 得到了SACC嗜神经侵袭PNI的相关差异基因,为阐明SACC的嗜神经侵袭机制提供了实验依据。

关键词: 唾液腺, 腺样囊性癌, 转录组测序技术, 差异表达基因, 嗜神经侵袭

Abstract: PURPOSE: To investigate the perineural invasion(PNI) related genes, the level of gene expression in single culture group and co-culture group of salivary adenoid cystic carcinoma (SACC) was analyzed by RNA-Seq technique. METHODS: Two groups of salivary adenoid cystic carcinoma (SACC) cells were analyzed for gene expression by co-culture system with rat Schwann cells (SCs). Then, cluster analysis, gene ontology (GO) function enrichment analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were used to analyze gene function, qRT-PCR was used to verify the 6 key-genes. Differential expression analysis of two conditions was performed using edgeR package (3.12.1). P value of 0.05 and log2 (fold change) of 1 were set as the threshold for significantly differential expression. RESULTS: The results showed that a total of 395 genes were differentially expressed in two groups (P≤0.05, |logFC|≥1.0), 135 genes were up-regulated (34.2%) and 260 genes were down-regulated (65.8%). GO enrichment analysis results showed that these PNI related DEGs involved in angiogenesis, extracellular matrix, cell proliferation, apoptosis and epithelial morphogenesis; KEGG pathway enrichment analysis displayed that these genes participated in histidine metabolism, interactions of tumor necrosis factor, chemotactic factor and cytokine receptor. qRT-PCR was used to validate 6 key-genes, and the results were consistent with RNA-Seq. CONCLUSIONS: Differentially expressed genes related to PNI of SACC were obtained, which provides important experimental basis for elucidating the mechanism of PNI of SACC, and also provides a new strategy and target for treatment of perineural invasion.

Key words: Salivary gland, Adenoid cystic carcinoma, Sequence analysis, RNA, DEGs, Perineural invasion

中图分类号:

R739.8

李欢, 杨新杰, 王维戚, 雷德林. 应用RNA-Seq技术筛选唾液腺腺样囊性癌嗜神经侵袭相关差异表达基因[J]. 中国口腔颌面外科杂志, 2018, 16(2): 114-119.

LI Huan, YANG Xin-jie, WANG Wei-qi, LEI De-lin. Primary study of the perineural invasion related genes of salivary adenoid cystic carcinoma by RNA-Seq[J]. China J Oral Maxillofac Surg, 2018, 16(2): 114-119.

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应用RNA-Seq技术筛选唾液腺腺样囊性癌嗜神经侵袭相关差异表达基因

Primary study of the perineural invasion related genes of salivary adenoid cystic carcinoma by RNA-Seq

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