目的 :探讨及分析细胞因子[白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和γ-干扰素(IFN-γ)]干预后小鼠胰岛细胞株MIN6中的长链非编码RNA(lnc RNA)表达谱的变化,并进行初步验证。方法:细胞因子干预小鼠胰岛细胞株MIN6,流式细胞仪检测细胞凋亡,CCK-8检测细胞活性,采用Affymetrix Gene Chip Mouse Transcriptome Array 1.0基因芯片筛查细胞因子干预组和正常对照组差异表达的lnc RNA,并采用实时荧光定量PCR对筛选出的部分lnc RNA进行验证。结果:与对照组相比,细胞因子干预组的细胞凋亡增加,细胞活性下降。细胞因子干预组中表达相差2倍以上的lnc RNA有723条,其中表达上调有444条,表达下调有279条。表达相差5倍以上的lnc RNA有111条,其中表达上调有105条,表达下调有6条。表达相差1.5倍以上的m RNA有2 180条。10条lnc RNA在细胞因子干预组中的表达经初步证实,结果与芯片趋势一致。结论:细胞因子干预后小鼠胰岛细胞株MIN6中lnc RNA的表达谱发生了显著变化,这些差异表达的lnc RNA在细胞因子诱导的胰岛细胞凋亡中具有重要作用。
Objective:To analyze lncRNA expression profiles in MIN6 cells,which belong to mouse pancreatic 13 cells, exposed to cytokines (IL-1β, TNF-α and IFN-γ) and to perform preliminary validation. Methods: MIN6 cells were incubated in the presence or absence of the cytokines mentioned aboved for 24 h. Cell apoptosis was detected by flow cytometry and cell viability was assessed using the Cell Counting kit-8. Affymetrix GeneChip Mouse Transcriptome Array 1.0 was performed to identify differently expressed lncRNA and mRNA in MIN6 cells exposed to cytokines compared with the normal control group. The interesting candidate lncRNAs were verified by Real-time quantitative PCR. Results: Cytokines promoted MIN6 cells apoptosis and inhibited the viability of MIN6 cells. A total of 723 differently expressed lncRNAs were identified in the cytokine-stimulated group compared with the control group with a set filter fold-change ≥ 2.0, of which 444 upregulated and 279 downregulated. Additionally, 111 differently expressed lncRNAs were identified with a set filter fold-change ≥ 5.0, of which 105 upregulated and 6 downregulated. And 2 180 differently expressed mRNAs were identified with a set filter fold-change ≥ 1.5. We also found, via quantitative PCR, that 10 lncRNAs were aberrantly expressed in the cytokine-stimulated group compared with the control group. The results of the qRT-PCR were consistent with the data from the microarray. Conclusion: The results revealed that there are differently expressed lncRNAs in MIN6 cells exposed to cytokines compared with the control group. These lncRNAs may play a key role in cytokine-induced apoptosis of pancreatic β cells.