lncRNA SNHG5对人类下咽癌细胞转录组学的影响及意义

doi:10.3969/j.issn.1673-8640.2022.03.018

检验医学 ›› 2022, Vol. 37 ›› Issue (3): 281-287.DOI: 10.3969/j.issn.1673-8640.2022.03.018

lncRNA SNHG5对人类下咽癌细胞转录组学的影响及意义

周焕¹, 唐颖², 杜翠萍², 杨扬², 冯会杰², 邱淼欣², 闫佩毅³, 蔡骁垚⁴, 金姝¹()

1.南华大学衡阳医学院,湖南衡阳 421001
2.上海市普陀区人民医院耳鼻喉科,上海 200060
3.上海市普陀区人民医院检验科,上海 200060
4.上海市普陀区人民医院中心实验室,上海 200060

收稿日期:2021-10-17 修回日期:2022-02-17 出版日期:2022-03-30 发布日期:2022-05-10
通讯作者: 金姝
作者简介:金姝,电子邮箱： jinshushu@sina.com。
周焕,男,1981年生,学士,主治医师,主要从事咽喉部肿瘤的基础研究。
基金资助:
上海市普陀区重点专科（耳鼻咽喉科）资助项目(2016zk105)

Effect of long non-coding RNA SNHG5 on the transcriptome of human hypopharyngeal carcinoma cells and clinical significance

ZHOU Huan¹, TANG Ying², DU Cuiping², YANG Yang², FENG Huijie², QIU Miaoxin², YAN Peiyi³, CAI Xiaoyao⁴, JIN Shu¹()

1. Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China
2. Department of Otolaryngology,Shanghai Putuo District People's Hospital,Shanghai 200060,China
3. Department of Clinical Laboratory,Shanghai Putuo District People's Hospital,Shanghai 200060,China
4. Department of Central Laboratory,Shanghai Putuo District People's Hospital,Shanghai 200060,China

Received:2021-10-17 Revised:2022-02-17 Online:2022-03-30 Published:2022-05-10
Contact: JIN Shu

摘要/Abstract

摘要：

目的探讨长链非编码RNA（lncRNA） SNHG5对人类下咽癌细胞株FaDu转录组学的影响。方法构建rLV-hSNHG5-ZsGreen-Puro慢病毒和rLV-ZsGreen-Puro对照慢病毒,将感染rLV-hSNHG5-ZsGreen-Puro慢病毒的FaDu细胞作为rLV-SNHG5组,将感染rLV-ZsGreen-Puro对照慢病毒的FaDu细胞作为rLV组。筛选差异基因集并进行基因转录能力、基因本体（GO）功能、京都基因与基因组数据库（KEGG）信号通路、Reactome通路、疾病本体（DO）功能分析。结果 rLV-SNHG5组SNHG5相对表达量显著高于rLV组（P<0.01）。基因差异分析共筛选出1 070个差异表达的基因,其中表达上调537个、表达下调533个。表达上调居前10位基因分别为LCP1、NT5E、CTGF、AXL、AKAP12、PXDN、THBS1、4-Mar、TMEM200A和RIPOR3,表达下调居前10位基因分别为PKP1、SERPINB13、KLK10、AC011473.4、NOTCH3、CA2、EGLN3、SLC6A8、KRT4和VAV3。功能分析结果显示,SNHG5改变了FaDu细胞部分基因的转录能力、参与调控GO功能、KEGG信号通路、Reactome通路和DO功能。结论 SNHG5对FaDu细胞的转录组学有一定的影响。

关键词: 小核仁RNA宿主基因5, 长链非编码RNA, 转录组学, 下咽癌

Abstract:

Objective To investigate the effect of long non-coding RNA（lncRNA） small nucleolar RNA host gene 5（SNHG5） on the transcriptome of human hypopharyngeal carcinoma cell line FaDu. Methods The rLV-hSNHG5-ZsGreen-Puro lentivirus and rLV-ZsGreen-Puro control lentivirus were constructed. FaDu cells infected with rLV-hSNHG5-ZsGreen-Puro lentivirus were served as rLV-SNHG5 group,and those infected with rLV-ZsGreen-Puro control lentivirus were served as rLV group. Differential gene sets were screened,and the gene transcriptional ability,Gene Ontology（GO） function,Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway,Reactome pathway and Disease Ontology（DO） function were analyzed. Results The relative expression of SNHG5 in rLV-SNHG5 group was higher than that in rLV group（P<0.01）. A total of 1 070 differentially expressed genes were screened by gene differential analysis,of which 537 were up-regulated,and 533 were down-regulated. The top 10 up-regulated genes with the most significant difference were LCP1,NT5E,CTGF,AXL,AKAP12,PXDN,THBS1,4-Mar,TMEM200A and RIPOR3,and the top 10 down-regulated genes were PKP1,SERPINB13,KLK10,AC011473.4,NOTCH3,CA2,EGLN3,SLC6A8,KRT4 and VAV3. Functional analysis showed that SNHG5 changed the gene transcriptional ability of some genes in FaDu cells and participated in the regulation of GO function,KEGG signaling pathway,Reactome pathway and DO function. Conclusions SNHG5 has a certain effect on the transcriptome of FaDu cells.

Key words: Small nucleolar RNA host gene 5, Long non-coding RNA, Transcriptomics, Hypopharyngeal carcinoma

中图分类号:

R393

周焕, 唐颖, 杜翠萍, 杨扬, 冯会杰, 邱淼欣, 闫佩毅, 蔡骁垚, 金姝. lncRNA SNHG5对人类下咽癌细胞转录组学的影响及意义[J]. 检验医学, 2022, 37(3): 281-287.

ZHOU Huan, TANG Ying, DU Cuiping, YANG Yang, FENG Huijie, QIU Miaoxin, YAN Peiyi, CAI Xiaoyao, JIN Shu. Effect of long non-coding RNA SNHG5 on the transcriptome of human hypopharyngeal carcinoma cells and clinical significance[J]. Laboratory Medicine, 2022, 37(3): 281-287.

图/表 9

图1 提取的RNA的琼脂糖凝胶电泳图

图2 rLV-SNHG5组与rLV组中SNHG5相对表达量的比较注：（a）RT-PCR扩增产物,M为Marker;（b）电泳条带灰度比值的比较。

图3 基因差异分析（a）注：差异基因火山图, 上调基因（537个）; 下调基因（533个）; （b）无差异基因（28 499个）;虚线表示差异基因筛选标准的阈值线;差异表达基因聚类热图;C1、C2、C3为rLV-SNHG5组样本;B1、B2、B3为rLV组样本;颜色越红,基因表达量越高;颜色越绿,基因表达量越低。

图4 rLV-SNHG5组与rLV组上调基因表达量的比较注：（a）LCP1;（b）NT5E;（c）CTGF;（d）AXL;（e）AKAP12;（f）PXDN;（g）THBS1;（h）4-Mar;（i）TMEM200A;（j）RIPOR3;与rLV组比较,*P<0.01。

图5 rLV-SNHG5组与rLV组下调基因表达量的比较注：（a）PKP1;（b）SERPINB13;（c）KLK10;（d）AC011473.4;（e）NOTCH3;（f）CA2;（g）EGLN3;（h）SLC6A8;（i）KRT4;（j）VAV3;与rLV组比较,*P<0.01。

图6 GO富集散点图注：（a）上调GO;（b）下调GO。

图7 KEGG富集分析注：（a）上调KEGG;（b）下调KEGG。

图8 Reactome通路分析注：（a）上调Reactome;（b）下调Reactome。

图9 DO功能分析注：（a）上调DO;（b）下调DO。

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lncRNA SNHG5对人类下咽癌细胞转录组学的影响及意义

Effect of long non-coding RNA SNHG5 on the transcriptome of human hypopharyngeal carcinoma cells and clinical significance

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