SIRT1调节人小梁网细胞中lncRNA相关信号通路的初步研究

Preliminary research on long non-coding RNA related signaling pathways regulated by SIRT1 in human trabecular meshwork cells

目的探讨沉默信息调节因子2相关酶1(SIRT1)调节人小梁网细胞功能的可能通路机制。方法实验研究。分别将SIRT1过表达慢病毒(SIRT1过表达组)和对照慢病毒(空载体对照组)按照最佳感染复数转染人小梁网细胞系,提取两组样本的总RNA进行长链非编码RNA(lncRNA)芯片检测。通过生物信息学技术对芯片差异性lncRNA进行分析,并用实时定量PCR法对筛选的差异性lncRNA进行验证。两组间比较采用独立样本t检验。结果lncRNA表达谱显示与空载体对照组相比,SIRT1过表达组人小梁网细胞有636个lncRNA上调和2246个lncRNA下调(差异倍数绝对值>2,均P<0.05)。基因本体分析显示SIRT1对人小梁网细胞外基质、细胞代谢、增殖、凋亡等有调节作用。京都基因与基因组百科全书生物学通路分析显示差异性lncRNA涉及19个信号通路,主要包括Notch信号通路、丝裂原激活蛋白激酶信号通路、酪氨酸酶相关蛋白通道的炎性反应介质调节以及细胞外基质受体的相互作用等。结论SIRT1能通过调节Notch、丝裂原激活蛋白激酶等多个信号通路的lncRNA影响小梁网细胞的功能。(中华眼科杂志,2021,57:215-222)

Objective To explore the possible mechanism of silent information regulator 1(SIRT1)in regulating the function of human trabecular meshwork cells(HTMCs).Methods Experimental study.HTMCs were transfected with SIRT1-overexpressed lentivirus and control lentivirus at the optimal multiplicity of infection,respectively.The total RNA was extracted,and the long non-coding RNA(lncRNA)microarray was used to detect the expression of lncRNAs.The differentially expressed lncRNAs were analyzed by bioinformatics.Real-time PCR was used to verify the microarray results.Student′s t-test was used for comparison between groups.Results Compared with the control group,there were 636 up-regulated lncRNAs and 2246 down-regulated lncRNAs in the SIRT1 overexpressed group(all P<0.05).Gene ontology analysis showed that SIRT1 regulated extracellular matrix,cell metabolism,proliferation and apoptosis of HTMCs.Kyoto encyclopedia of genes and genomics pathway analysis indicated that differential lncRNAs induced by SIRT1 were involved in 19 signal pathways,including Notch signaling pathway,mitogen-activated protein kinase signaling pathway,inflammatory mediator regulation of tyrosinase-associated protein channels and extracellular matrix-receptor interaction.Conclusion SIRT1 could have effects on the function of HTMCs through regulating lncRNAs in multiple signaling pathways,including Notch signaling pathway,mitogen-activated protein kinase signaling pathway,etc.(Chin J Ophthalmol,2021,57:215-222)