PLAG1转基因小鼠中差异表达长链非编码RNA的筛选分析

Study on the differentially expressed long non-coding RNAs in PLAG1 transgenic mice by gene micro-array

目的:应用基因芯片技术,筛选PLAG1转基因小鼠下颌下区多形性腺瘤中差异表达的长链非编码RNA(LncRNA)。方法:取3对PLAG1转基因小鼠下颌下区多形性腺瘤组织、对侧无瘤腺体组织、空白对照小鼠腺体组织,抽提标本总RNA并反转录合成cDNA探针,与芯片杂交后获得荧光信号,分析在肿瘤组织中差异表达的长链非编码RNA。随机挑选其中5个LncRNA,利用实时荧光定量PCR在6对样本中验证其表达。采用SPSS13.0软件包中的t检验评价表达差异。结果:基因芯片结果显示,与PLAG1转基因小鼠无瘤腺体组织、空白对照小鼠腺体组织相比,PLAG1转基因小鼠肿瘤中共有5627个LncRNA同时发生差异表达,其中2871个上调、2756个下调。实时荧光定量PCR结果显示,5个随机挑选的LncRNA中,4个(AK146794、ENSMUST00000119440、ENSMUST00000140495、Gm12873)与基因芯片结果一致。结论:应用基因芯片筛选出PLAG1转基因小鼠中差异表达的长链非编码RNA,为进一步研究多形性腺瘤的发病机制提供了依据。

PURPOSE: To screen for the differentially expressed long non-coding RNAs(LncRNAs) in pleomorphic adenoma of PLAG1 transgenic mice. METHODS: Three pairs of pleomorphic adenoma, non-tumor gland tissues from PLAG1 transgenic mice and normal gland tissues of submandibular gland from wide-type mice were collected and then the total RNA was extracted. cDNA probe was synthesized by reverse transcription and hybridized as the cDNA microarray for scanning fluorescent signals and differentially expressed LncRNAs. The expressions of 5 randomly selected LncRNAs were validated in another 6 pairs of tissues via real-time PCR. The expression differences were compared and analyzed using SPSS13.0 software package. RESULTS: Microarray results demonstrated that a total of 5627 LncRNAs(2871 upregulated and 2756 downregulated) were differentially expressed in the tumors of PLAG1 transgenic mice. Real-time PCR data showed that the expression of 4 selected LncRNAs(AK146794, ENSMUST00000119440, ENSMUST00000140495 and Gm12873) were consistent with the microarray results. CONCLUSIONS: Analysis of differentially expressed LncRNAs in PLAG1 transgenic mice can provide evidence for pathogenesis of pleomorphic adenoma.