目的:阐明反流性食管炎中差异表达的基因并探讨其造成反流性食管炎黏膜损伤的可能机制,初步筛选并鉴定长链非编码RNA HOTAIR在反流性食管炎中的表达变化和作用。方法:取内镜下反流性食管炎黏膜糜烂处组织标本及糜烂旁黏膜光滑食管组织进行转录组测序和生物信息学分析,实时荧光定量PCR测定两类组织中长链非编码RNA HOTAIR的表达变化,并分析其与反流性食管炎患者年龄的关系。结果:转录组测序结果显示反流性食管炎糜烂组和糜烂旁组之间有多种基因差异表达,其中7 661个基因表达下调。生物信息学分析显示差异表达的基因富集于免疫反应。对于筛选出的长链非编码RNA HOTAIR,实时荧光定量PCR显示其相对表达量与反流性食管炎患者年龄相关(P＜0.05)。相较于糜烂旁组,大年龄组中反流性食管炎糜烂组的HOTAIR表达上调(P＜0.05),而在小年龄组中反流性食管炎糜烂组HOTAIR表达下调(P＜0.05)。结论:反流性食管炎多种差异表达的基因可能通过免疫反应参与该病的发生发展过程。筛选出的长链非编码RNA HOTAIR表达变化在不同年龄患者中呈现相反的趋势,提示HOTAIR可能在反流性食管炎中发挥重要作用并可作为诊断的重要辅助指标。
Objective: To elucidate the differentially expressed genes in reflux esophagitis and explore the possible mechanism of mucosal damage in reflux esophagitis, and preliminarily screen and identify the expression changes and effects of long-chain non-coding RNA HOTAIR in reflux esophagitis. Methods: Transcriptome sequencing and bioinformatics analysis were performed on tissue specimens at mucosal erosion site of reflux esophagitis and mucosa smooth esophageal tissue under para-erosion endoscopy. The expression changes of long-chain non-coding RNA HOTAIR in the two types of tissues were determined by real-time fluorescent quantitative PCR, and its relationship with the age of patients with reflux esophagitis was analyzed. Results: The transcriptome sequencing results showed that there were a variety of genes differentially expressed between the erosive group and the para-erosive group of reflux esophagitis, of which 7661 genes were down-regulated. Bioinformatics analysis showed that differentially expressed genes were enriched in immune responses. For the screened long-chain non-coding RNA HOTAIR, real-time fluorescent quantitative PCR showed that its relative expression was correlated with the age of patients with reflux esophagitis (P＜0.05). Compared with the para-erosion group, the expression of HOTAIR was up-regulated in the erosion group of reflux esophagitis in the older age group (P＜0.05), and down-regulated in the erosion group of reflux esophagitis in the younger age group (P＜0.05). Conclusion: The differentially expressed genes may be involved in the pathogenesis of reflux esophagitis through immune response. The screened long-chain non-coding RNA HOTAIR expression showed opposite trends in patients of different ages, suggesting that HOTAIR may play an important role in reflux esophagitis and can be used as an important auxiliary indicator for diagnosis.
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