| 摘要: |
| 目的 探讨长链非编码RNA(lncRNA)在βB2基因敲除小鼠睾丸中的表达及其影响睾丸发育的可能机制。 方法 采用lncRNA芯片技术,筛选野生型(WT)和βB2基因敲除(KO)小鼠睾丸组织(均n=3)中lncRNA及信使RNA(mRNA)表达谱的变化;对差异表达lncRNA和mRNA进行GO数据库分析及KEGG数据库分析,建立调控网络图;采用实时荧光定量PCR(qRT-PCR)验证差异表达的lncRNA和mRNA,探讨调控通路。 结果 (1)两组小鼠睾丸组织差异表达的lncRNA共140条,mRNA共477条;(2)通过GO数据库分析,筛选出差异表达lncRNA共12条,其中上调7条,下调5条;(3)经KEGG数据库进行路径分析,发现差异表达mRNA主要经由Ca2+信号、配体-受体相互作用等信号通路起作用;(4)用关联矩阵法建立了lncRNA和mRNA共表达网络图,共有17个节点,12条连接,包含9条lncRNA和8条mRNA;其中Rsl1由3条lncRNA调控,Lpo和Mpo各由2条lncRNA调控,Hdac1、Ephb4等各由1条lncRNA调控。(5)qRT-PCR分析结果表明,在βB2基因敲除小鼠睾丸组织中,lncRNA A-30-P01019163和P2rx7的表达均下调(P<0.05)。 结论 lncRNA与βB2基因的晶状体外功能密切相关,其中lncRNA A-30-P01019163可能通过调控下游P2rx7 mRNA的表达影响睾丸组织细胞周期及信号转导,进而调控睾丸发育。 |
| 关键词: lncRNA βB2晶体蛋白 睾丸发育 P2rx7 |
| DOI：10.16781/j.0258-879x.2016.01.0059 |
| 投稿时间：2015-10-12修订日期：2015-12-05 |
| 基金项目:国家自然科学基金(81170834, 81571387, 81300748). |
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| LncRNAs expression profile in testis tissues of CRYBB2 gene knockout mice |
| REN Han-xiao1△,GAO Qian1△,JIA Yin1,ZHANG Jian-rong1,ZHANG Jun-jie2*,LI Wen-jie1* |
(1. Department of Laboratory Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Obstetrics and Gynecology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
△Co-first authors.
*Corresponding authors.) |
| Abstract: |
| Objective To explore the expression profile of lncRNAs in the testis tissue of CRYBB2 gene knockout (KO) mice and its possible role in the testis development. Methods Testis tissues(n=3)from wild-type (WT) and CRYBB2 KO mice were subjected to lncRNA and mRNA microarray profiling. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to group the differentially expressed lncRNAs into regulated gene pathways and functions. The correlation matrix method was used to establish a network of lncRNA and mRNA co-expression. Quantitative (q)RT-PCR was used to verify expression of some differentially expressed lncRNAs and mRNAs. Results There were 140 differentially expressed lncRNAs and 477 differentially expressed mRNAs between testis tissues from WT and KO mice. There were 12 differentially expressed lncRNAs through the analyses of the GO, with 7 up-regulated and 5 down-regulated. The KEGG analysis showed that these differentially expressed mRNAs played important roles in Ca2+ signaling, ligand and receptor interactions, and so on. The correlation matrix method established an lncRNA and mRNA co-expression network, consisting of 9 lncRNAs and 8 mRNAs, with 17 nodes and 12 connections. Furthermore, expression of gene Rsl1 was regulated by three lncRNAs, expression of gene Lpo and gene Mpo was regulated by two lncRNAs, and expression of gene Hdac1 and gene Ephb4 was regulated by one lncRNA. qRT-PCR confirmed the significant down-regulation of lncRNA A-30-P01019163 expression, which significantly down-regulated its downstream gene P2rx7 in testis tissues of CRYBB2 KO mice(P<0.05). Conclusion LncRNA is closely related to the non-crystalline lens function of CRYBB2. LncRNA A-30-P01019163 may affect testicular cell cycle and signaling pathway by regulating P2rx7 expression in the testis tissues. |
| Key words: lncRNA CRYBB2 testis development P2rx7 |
