寻常型天疱疮外周血长链非编码RNA的差异性表达
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摘要
目的筛选寻常型天疱疮(PV)患者外周血单个核细胞(PBMC)和正常人PBMC中差异性表达的长链非编码RNA(LncRNA)。方法利用LncRNA表达谱芯片检测5例PV和3例正常人PBMC中LncRNA的表达,以P<0.05,且差异表达倍数大于2或小于0.5的条件筛选出差异性表达的LncRNA,并采用Real-time PCR进一步验证其在40例PV患者和40例正常人PBMC中的表达情况。结果与正常人相比,PV患者PBMC中差异性表达的LncRNA共有219个,其中上调142个,下调77个。选取的4个LncRNA在40例PV患者和40例正常人PBMC中的验证结果为:Lnc-BST1-1∶1,NR_023343,Lnc-ERICH1-9∶3表达上调(P<0.05),Lnc-CABIN1-1∶1表达下调(P<0.05)。与基因芯片检测结果相符。结论多个长链非编码RNA在PV患者外周血单个核细胞中异常表达,提示它们可能在PV的发生发展中起着重要的调控作用。
Objective To elucidate the differential expression of long-chain non-coding RNA(LncRNA) in peripheral blood mononuclear cells(PBMC) in patients with pemphigus vulgaris(PV) and normal human PBMC.Methods LncRNA expression profiling microarray was used to detect the expression of LncRNA in 5 cases of PV and 3 cases of normal human PBMCs.Differential expression of LncRNA was screened for those with significance of P<0.05)and differences ≥2 or≤0.5 folds.Real-time PCR was used to further verify its expression in 40 cases of PV patients and 40 cases of normal PBMC.Results Compared with normal people, there were 219 LncRNAs differentially expressed in PBMCs of PV patients, including 142 up-regulated and 77 down-regulated.The validation of the four selected LncRNAs in 40 PV patients and 40 normal human PBMCs demonstrated upregulations of Lnc-BST1-1∶1, NR_023343 and Lnc-ERICH1-9∶3 expression(P<0.05), and down-regulation of lnc-CABIN1-1∶1(P<0.05).This results were consistent with gene chip test results.Conclusion There is a differential expression of LncRNA in PBMCs in PV patients compared with normal controls, indicating its important role in the development of PV.
Objective To elucidate the differential expression of long-chain non-coding RNA(LncRNA) in peripheral blood mononuclear cells(PBMC) in patients with pemphigus vulgaris(PV) and normal human PBMC.Methods LncRNA expression profiling microarray was used to detect the expression of LncRNA in 5 cases of PV and 3 cases of normal human PBMCs.Differential expression of LncRNA was screened for those with significance of P<0.05)and differences ≥2 or≤0.5 folds.Real-time PCR was used to further verify its expression in 40 cases of PV patients and 40 cases of normal PBMC.Results Compared with normal people, there were 219 LncRNAs differentially expressed in PBMCs of PV patients, including 142 up-regulated and 77 down-regulated.The validation of the four selected LncRNAs in 40 PV patients and 40 normal human PBMCs demonstrated upregulations of Lnc-BST1-1∶1, NR_023343 and Lnc-ERICH1-9∶3 expression(P<0.05), and down-regulation of lnc-CABIN1-1∶1(P<0.05).This results were consistent with gene chip test results.Conclusion There is a differential expression of LncRNA in PBMCs in PV patients compared with normal controls, indicating its important role in the development of PV.
引文
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[2] Dick SE,Werth VP.Pemphigus:A treatment update [J].Autoimmunity,2006,39(7):591-599.
[3] Anhalt GJ,Diaz LA.Prospects for autoimmune disease:Research advances inpemphigus[J].JAMA,2001,285(6):652-654.
[4] Ikeda S,Imamura S,Hashimoto I,et al.History of the establishment andrevision of diagnostic criteria,severity index and therapeutic guidelines forpemphigus in Japan[J].Arch Dermatol Res,2003,295(Suppl 1):S12-S16.
[5] Meyer N,Misery L.Geoepidemiologic considerations of auto-immunepemphigus[J].Autoimmun Rev,2010,9(5):A379-382.
[6] Ahmed AR,Moy R.Death in pemphigus[J].J Am Acad Dermatol,1982,7(2):221-228.
[7] 王慧丛,付萍.CD4+T淋巴细胞与寻常型天疱疮相关性研究进展[J].皮肤病与性病,2014,36(1):17-19 .
[8] 尚建基.天疱疮58例临床分析[J].中国中西医结合皮肤性病学杂志,2007,27(3):159 .
[9] Sarig O,Bercovici S,Zoller L,et al.Population-specific association between a polymorphic variant in ST18,encoding a pro-apoptotic molecule,and pemphigus vulgaris[J].J Invest Dermatol,2012,132(7):1798-1805.
[10] Qrom UA,Derrien T,Beringer M,et al.Long noncoding RNAs with enhancer-like function in human cells[J].Cell,2010,143(1):46-58.
[11] Zhou J,Yang L,Zhong T,et al.H19 lncRNA alters DNA methylationgenome wide byregulating S-adenosylhomocysteine hydrolase[J].Nat Commun,2015,21(6):10221.
[12] Huang W,Thomas B,Flynn RA,et al.DDX5 andits associated lncRNARmrp modulate TH17 cell effector functions[J].Nature,2015,528(7583):517-522.
[13] Chan AS,Thorner PS,Squire JA,et al.Identification of a novelgene NCRMS on chromosome 12q21 with differential expression betweenrhabdomyosarcoma subtypes[J].Oncogene,2002,21(19):3029-3037.
[14] Mercer TR,Dinger ME,Mattick JS.Long non-coding RNAs:insights intofunctions[J].Nat Rev Genet,2009,10(3):155-159.
[15] Szegedi K,Sonkoly E,Nagy N,et al.The anti-apoptotic protein G1P3 is overexpressed inpsoriasis and regulated by the non-coding RNA,PRINS[J].Exp Dermatol,2010,19(3):269-278.
[16] Ishii N,Ozaki K,Sato H,et al.Identification of a novel non-coding RNA,MIAT,that confers risk of myocardial infarction[J].J Hum Genet,2006,51(12):1087-1099.
[17] Wang K,Liu CY,Zhou LY,et al.APF lncRNA regulates autophagy andmyocardial infarction by targeting miR-188-3p[J].Nat Commun,2015,33(3):245-249.
[18] 生物通.lncRNA:一个全新的世界[EB/OL].(2013-07-03)http://www.ebiotrade.com/newsf/2013-7/201373 100128779.htm.