小鼠颌骨发育过程中DMP1基因相关lncRNA的表达研究
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摘要
目的研究C57小鼠不同发育时期颌骨中DMP1基因附近长链非编码RNA (lncRNA)的表达变化情况。方法利用lncRNA测序技术和实时定量PCR (RT-PCR)检测孕18 d (E18D)及出生后2周(2W)的C57小鼠下颌骨组织样本中DMP1基因位置附近的lncRNA表达变化,分析其与DMP1基因表达变化的关系。结果 E18D组与2W组间有6 617条差异表达的lncRNA (∣log2 (2W/E18D)∣>1且控制错误发现率<0.01),其中有5条lncRNA在DMP1基因上游或下游300 000 bp以内。RT-PCR分析显示,2W组中DMP1和lnc19450的表达较E18D组降低,而lnc30219、lnc8292、lnc30219和lnc32865的表达较E18D组上升(P均<0.05)。结论 lnc19450的表达与DMP1基因的表达趋势一致,lnc30219、lnc8292、lnc30219和lnc32865的表达与DMP1基因的表达趋势相反,推测lnc19450、 lnc30219、 lnc8292、lnc30219和lnc32865可能在DMP1基因的表达调控起着重要的作用。
Objective To analyze the changes in the expression of long non-coding RNA(1ncRNA) adjacent to the DMP1 gene in different developmental stages of jawbone in C57 mice.Methods The lncRNA sequencing and quantitative real-time PCR(RT-PCR) were performed to detect the expression profile of DMP1 gene-related lncRNAs at the embryonic day 18(E18D) and 2 weeks after birth(2W) in the jawbone of C57 mice,and analyze the relationship between lncRNA expression and the changes in the expression of DMP1.Results Compared with the E18D group,6 617 lncRNAs were differentially expressed in the 2 W group(∣log2(2W/E18D) ∣>1 and FDR<0.01),among which,5 lncRNAs were at the upstream or downstream of DMP1 gene position within 300 kb. RT-PCRdemonstrated that the expression levels of DMP1 and lnc19450 in the 2 W group were lower than those in the E18D group, whereas the expression levels of lnc30219, lnc8292,lnc30219 and lnc32865 were higher compared with those in the E18 D group(all P<0.05).Conclusions The expression trends of lnc19450 and DMP1 are consistent, whereas the expression patterns of lnc30219,lnc8292,lnc30219 and lnc32865 are opposite with that of DMP1 gene,suggesting that lnc19450,lnc30219,lnc8292,lnc30219 and lnc32865 may play a vital role in regulating the expression of DMP1 gene.
Objective To analyze the changes in the expression of long non-coding RNA(1ncRNA) adjacent to the DMP1 gene in different developmental stages of jawbone in C57 mice.Methods The lncRNA sequencing and quantitative real-time PCR(RT-PCR) were performed to detect the expression profile of DMP1 gene-related lncRNAs at the embryonic day 18(E18D) and 2 weeks after birth(2W) in the jawbone of C57 mice,and analyze the relationship between lncRNA expression and the changes in the expression of DMP1.Results Compared with the E18D group,6 617 lncRNAs were differentially expressed in the 2 W group(∣log2(2W/E18D) ∣>1 and FDR<0.01),among which,5 lncRNAs were at the upstream or downstream of DMP1 gene position within 300 kb. RT-PCRdemonstrated that the expression levels of DMP1 and lnc19450 in the 2 W group were lower than those in the E18D group, whereas the expression levels of lnc30219, lnc8292,lnc30219 and lnc32865 were higher compared with those in the E18 D group(all P<0.05).Conclusions The expression trends of lnc19450 and DMP1 are consistent, whereas the expression patterns of lnc30219,lnc8292,lnc30219 and lnc32865 are opposite with that of DMP1 gene,suggesting that lnc19450,lnc30219,lnc8292,lnc30219 and lnc32865 may play a vital role in regulating the expression of DMP1 gene.
引文
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[20]Kamiya N,Takagi M.Differential expression of dentin matrix protein 1,type I collagen and osteocalcin genes in rat developing mandibular bone.Histochem J,2001,33(9-10):545-552.
[2]Tuo YL,Li XM,Luo J.Long noncoding RNA UCA1 modulates breast cancer cell growth and apoptosis through decreasing tumor suppressive miR-143.Eur Rev Med Pharmacol Sci,2015,19(18):3403-3411.
[3]Zhao W,An Y,Liang Y,Xie XW.Role of HOTAIR long noncoding RNA in metastatic progression of lung cancer.Eur Rev Med Pharmacol Sci,2014,18(13):1930-1936.
[4]Marchese FP,Raimondi I,Huarte M.The multidimensional mechanisms of long noncoding RNA function.Genome Biol,2017,18(1):206.
[5]Long Y,Wang X,Youmans DT,Cech TR.How do lncRNAs regulate transcription?Sci Adv,2017,3(9):eaao2110.
[6]Ravindran S,George A.Dentin matrix proteins in bone tissue engineering.Adv Exp Med Biol,2015,881:129-142.
[7]Jing B,Zhang C,Liu X,Zhou L,Liu J,Yao Y,Yu J,Weng Y,Pan M,Liu J,Wang Z,Sun Y,Sun YE.Glycosylation of dentin matrix protein 1 is a novel key element for astrocyte maturation and BBB integrity.Protein Cell,2018,9(3):298-309.
[8]Sun Y,Lu Y,Chen L,Gao T,D'Souza R,Feng JQ,Qin C.DMP1 processing is essential to dentin and jaw formation.J Dent Res,2011,90(5):619-624.
[9]Liao Q,Liu C,Yuan X,Kang S,Miao R,Xiao H,Zhao G,Luo H,Bu D,Zhao H,Skogerb?G,Wu Z,Zhao Y.Largescale prediction of long non-coding RNA functions in a coding-non-coding gene co-expression network.Nucleic Acids Res,2011,39(9):3864-3878.
[10]邓鹏辉,黄传君,赵方正,宫晓丹,李春晓,张才擎.TGF-β1、VEGF在哮喘小鼠的表达及细辛脑的干预作用.新医学,2018,49(1):25-29.
[11]Ponting CP,Oliver PL,Reik W.Evolution and functions of long noncoding RNAs.Cell,2009,136(4):629-641.
[12]Schmitz SU,Grote P,Herrmann BG.Mechanisms of long noncoding RNA function in development and disease.Cell Mol Life Sci,2016,73(13):2491-2509.
[13]Spadaro PA,Flavell CR,Widagdo J,Ratnu VS,Troup M,Ragan C,Mattick JS,Bredy TW.Long noncoding RNA-directed epigenetic regulation of gene expression is associated with anxietylike behavior in mice.Biol Psychiatry,2015,78(12):848-859.
[14]Terashima M,Tange S,Ishimura A,Suzuki T.MEG3 long noncoding RNA contributes to the epigenetic regulation of epithelialmesenchymal transition in lung cancer cell lines.J Biol Chem,2017,292(1):82-99.
[15]Wang Y,Zhong H,Xie X,Chen CY,Huang D,Shen L,Zhang H,Chen ZW,Zeng G.Long noncoding RNA derived from CD244 signaling epigenetically controls CD8+T-cell immune responses in tuberculosis infection.Proc Natl Acad Sci U S A,2015,112(29):E3883-E3892.
[16]Ding G,Li W,Liu J,Zeng Y,Mao C,Kang Y,Shang J.LncRNA GHET1 activated by H3K27 acetylation promotes cell tumorigenesis through regulating ATF1 in hepatocellular carcinoma.Biomed Pharmacother,2017,94:326-331.
[17]Thotakura SR,Karthikeyan N,Smith T,Liu K,George A.Cloning and characterization of rat dentin matrix protein 1(DMP1)gene and its 5'-upstream region.J Biol Chem,2000,275(14):10272-10277.
[18]Chen S,Chen L,Jahangiri A,Chen B,Wu Y,Chuang HH,Qin C,MacDougall M.Expression and processing of small integrin-binding ligand N-linked glycoproteins in mouse odontoblastic cells.Arch Oral Biol,2008,53(9):879-889.
[19]Mashhadi Abbas F,Sichani Fallahi H,Khoshzaban A,Mahdavi N,Bagheri SS.Expression of odontogenic genes in human bone marrow mesenchymal stem cells.Cell J,2013,15(2):136-141.
[20]Kamiya N,Takagi M.Differential expression of dentin matrix protein 1,type I collagen and osteocalcin genes in rat developing mandibular bone.Histochem J,2001,33(9-10):545-552.