基于RNA测序研究过表达AJUBA对T47D基因表达谱的影响

doi:10.3969/j.issn.1673-8640.2016.06.018

检验医学 ›› 2016, Vol. 31 ›› Issue (6): 527-532.DOI: 10.3969/j.issn.1673-8640.2016.06.018

基于RNA测序研究过表达AJUBA对T47D基因表达谱的影响

徐北惠¹, 李琪², 邹秀群², 徐洪², 侯照远², 王家敏², 倪培华¹

1.上海交通大学医学院附属瑞金医院检验系,上海 200025
2.上海交通大学医学院生物化学与分子细胞生物学系,上海 200025

收稿日期:2016-02-22 出版日期:2016-06-30 发布日期:2016-07-05
作者简介:null
作者简介：徐北惠,女,1989年生,硕士,主要从事肿瘤发生、发展的机制研究。

通讯作者：倪培华,联系电话：021-64370045-610505。

Influence of AJUBA overexpression on T47D gene expression profile by RNA sequencing

XU Beihui¹, LI Qi², ZOU Xiuqun², XU Hong², HOU Zhaoyuan², WANG Jiamin², NI Peihua¹

1. Department of Laboratory Medicine,Ruijin Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China
2. Department of Biochemistry and Molecular Cytobiology,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China

Received:2016-02-22 Online:2016-06-30 Published:2016-07-05

摘要/Abstract

摘要：

目的探究AJUBA基因对于雌激素受体阳性乳腺癌细胞T47D基因表达的影响。方法构建AJUBA稳定过表达的T47D细胞系。载体对照组和实验组分别提取RNA进行转录组测序技术（RNA-seq）测序,将所得序列映射到人类基因组并进行转录组重建,样本标准化后寻找实验组和对照组之间的差异基因,利用生物信息学方法进一步对所得的差异基因进行基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）通路富集性分析,同时挑选部分基因用实时荧光定量聚合酶链反应（RT-qPCR）进行基因表达验证。结果实验组与对照组细胞相比共找到568个差异基因,其中上调基因239个,下调基因329个。GO分析中,注释到分子功能、生物学过程和细胞组成的上调差异基因分别有3、23、8个;下调差异基因分别有21、35、9个。KEGG分析中上调基因显著富集通路有2个,下调基因显著富集通路有4个。结论 AJUBA过表达可以影响T47D细胞的一系列生物学过程相关的多条信号通路,可能在乳腺癌发生、发展中起重要作用。

关键词: AJUBA, 乳腺癌, 高通量RNA测序, 差异表达基因, 生物信息学分析

Abstract:

Objective To investigate the influence of AJUBA gene on estrogen receptor-positive breast cancer cell line T47D gene expression. Methods T47D cell line for stable expression of AJUBA was established. RNA-seq analysis was performed for RNA extracted from T47D-vector and T47D-AJUBA groups. After mapping and reading,transcriptome reconstruction,normalization and expression quantification,differentially expressed genes were identified with gene ontology（GO） analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis. Differentially expressed genes were validated by real-time fluorescence quantitative polymerase chain reaction（RT-qPCR）. Results A total of 568 differentially expressed genes were identified,including 239 up-regulated genes and 329 down-regulated genes. In GO analysis,3,23 and 8 up-regulated genes were annotated to molecular function,biological process and cellular component,respectively,while 21,35 and 9 down-regulated genes were also annotated to molecular function,biological process and cellular component,respectively. In KEGG analysis,2 were up-regulated genes,and 4 were down-regulated genes. Conclusions The overexpression of AJUBA gene may have remarkable effects on a series of biological function-related multiple signal pathways,and AJUBA gene might play an important role in the tumorigenesis and development of breast cancer.

Key words: AJUBA, Breast cancer, High-throughput RNA sequencing, Differentially expressed gene, Bioinformatics analysis

中图分类号:

R446.1

徐北惠, 李琪, 邹秀群, 徐洪, 侯照远, 王家敏, 倪培华. 基于RNA测序研究过表达AJUBA对T47D基因表达谱的影响[J]. 检验医学, 2016, 31(6): 527-532.

XU Beihui, LI Qi, ZOU Xiuqun, XU Hong, HOU Zhaoyuan, WANG Jiamin, NI Peihua. Influence of AJUBA overexpression on T47D gene expression profile by RNA sequencing[J]. Laboratory Medicine, 2016, 31(6): 527-532.

图/表 4

参考文献 17

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作用	途径	基因数	占总基因百分比（%）	P值
上调	PPAR 信号通路	4	2.515 723 27	0.012 399 522
	ErbB 信号通路	4	2.515 723 27	0.023 003 129
下调	轴突导向	7	2.857 142 857	0.005 757 691
	VEGF信号通路	5	2.040 816 327	0.015 001 19
	肥厚性心肌病	5	2.040 816 327	0.022 738 371
	扩张性心肌病	5	2.040 816 327	0.029 377 142

作用	途径	基因数	占总基因百分比（%）	P值
上调	PPAR 信号通路	4	2.515 723 27	0.012 399 522
	ErbB 信号通路	4	2.515 723 27	0.023 003 129
下调	轴突导向	7	2.857 142 857	0.005 757 691
	VEGF信号通路	5	2.040 816 327	0.015 001 19
	肥厚性心肌病	5	2.040 816 327	0.022 738 371
	扩张性心肌病	5	2.040 816 327	0.029 377 142

基于RNA测序研究过表达AJUBA对T47D基因表达谱的影响

Influence of AJUBA overexpression on T47D gene expression profile by RNA sequencing

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