熊新威,郑哲,焦钰,邓岳文,杜晓东

• 1:广东海洋大学水产学院广东省珍珠养殖与加工工程技术研究中心

摘要(Abstract)：

用马氏珠母贝基因组序列,通过生物信息学方法获得Pm-miR-183的前体,长度76 bp,具有典型的茎环结构。成熟序列及前体序列的同源性分析均显示较高的种间保守性。Pm-miR-183前体序列(Pm-pre-miR-183)的系统进化树分析,Pm-pre-miR-183与帽贝同源性较高。靶向关系分析,Pm-miR-183与矿化基因及免疫相关基因均存在靶向作用位点。实时荧光定量分析,Pm-miR-183在马氏珠母贝的各组织中均有表达。注射Pm-miR-183mimics进行过表达,Pm-miR-183的表达量在套膜区(MP)中显著上调(P<0.05)。扫描电子显微镜(scanning electron microscope,SEM)检测,Pm-miR-183过表达会导致贝壳珍珠层生长出现紊乱。

关键词(KeyWords)： 马氏珠母贝;Pm-miR-183;生物矿化;miRNA调控

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31672626);; 广东省海洋渔业科技推广专项(B201601-Z10)

作者(Author): 熊新威,郑哲,焦钰,邓岳文,杜晓东

参考文献(References)：

• [1]XU F,WANG X T,FENG Y,et al.Identification of conserved and novel micro RNAs in the Pacific oyster Crassostrea gigas by deep sequencing[J].PLo S One,2014,9(8):e104371.
• [2]LEWIS B P,BURGE C B,BARTEL D P.Conserved seed pairing,often flanked by adenosines,indicates that thousands of human genes are Micro RNA targets[J].Cell,2005,120(1):15-20.
• [3]LIM L P,LAU N C,GARRETT-ENGELE P,et al.Microarray analysis shows that some micro RNAs downregulate large numbers of target m RNAs[J].Nature,2005,433(7027):769-773.
• [4]潘晓艳,郑哲,田荣荣,等.mi R-210在马氏珠母贝外套膜矿化机制中的结构和功能预测[J].广东海洋大学学报,2016,36(3):9-14.
• [5]KAPINAS K,DELANY A M.Micro RNA biogenesis and regulation of bone remodeling[J].Arthritis Research&Therapy,2011,13(3):220.
• [6]ZHAO X,XU D,LI Y,et al.Micro RNAs regulate bone metabolism[J].Journal of Bone and Mineral Metabolism,2014,32(3):221-231.
• [7]崔福斋.生物矿化[M].2版.北京:清华大学出版社,2012.
• [8]TIAN R R,ZHENG Z,HUANG R L,et al.mi R-29a participated in nacre formation and immune response by targeting Y2R in Pinctada martensii[J].International Journal of Molecular Sciences,2015,16(12):29436-29445.
• [9]JIAO Y,ZHENG Z,TIAN R R,et al.Micro RNA,Pm-mi R-2305,participates in nacre formation by targeting pearlin in pearl oyster Pinctada martensii[J].International Journal of Molecular Sciences,2015,16(9):21442-21453.
• [10]ZHENG Z,JIAO Y,DU X,et al.Computational prediction of candidate mi RNAs and their potential functions in biomineralization in pearl oyster Pinctada martensii[J].Saudi Journal of Biological Sciences,2016,23(3):372-378.
• [11]PIERCE M L,WESTON M D,FRITZSCH B,et al.Micro RNA-183 family conservation and ciliated neurosensory organ expression[J].Evolution&Development,2008,10(1):106-113.
• [12]LOWERY A J,MILLER N,DWYER R M,et al.Dysregulated mi R-183 inhibits migration in breast cancer cells[J].BMC Cancer,2010,10:502.
• [13]ZHU J F,FENG Y P,KE Z F,et al.Down-regulation of mi R-183 promotes migration and invasion of osteosarcoma by targeting Ezrin[J].The American Journal of Pathology,2012,180(6):2440-2451.
• [14]KUNDU S T,BYERS L A,PENG D H,et al.The mi R-200 family and the mi R-183~96~182 cluster target Foxf2 to inhibit invasion and metastasis in lung cancers[J].Oncogene,2016,35(2):173-186.
• [15]CHENG Y,XIANG G X,MENG Y B,et al.Mi RNA-183-5p promotes cell proliferation and inhibits apoptosis in human breast cancer by targeting the PDCD4[J].Reproductive Biology,2016,16(3):225-233.
• [16]KLOOSTERMAN W P,WIENHOLDS E,DE BRUIJN E,et al.In situ detection of mi RNAs in animal embryos using LNA-modified oligonucleotide probes[J].Nature Methods,2006,3(1):27-29.
• [17]张群.文昌鱼mi R-183和mi R-216基因簇的表达模式和保守性研究[D].青岛:中国海洋大学,2012.
• [18]VAN DEN ACKERVEKEN P,MOUNIER A,HUYGHE A,et al.The mi R-183/Itg A3 axis is a key regulator of prosensory area during early inner ear development[J].Cell Death and Differentiation,2017,24:2054-2065.
• [19]LIU W H,CHANG L S.Suppression of Akt/Foxp3-mediated mi R-183 expression blocks Sp1-mediated ADAM17 expression and TNFα-mediated NFκB activation in piceatannol-treated human leukemia U937 cells[J].Biochemical Pharmacology,2012,84(5):670-680.
• [20]SHI X Y,GU L,CHEN J,et al.Downregulation of mi R-183 inhibits apoptosis and enhances the invasive potential of endometrial stromal cells in endometriosis[J].International Journal of Molecular Medicine,2014,33(1):59-67.
• [21]DONATELLI S S,ZHOU J M,GILVARY D L,et al.TGF-β-inducible micro RNA-183 silences tumor-associated natural killer cells[J].Proceedings of the National Academy of Sciences of the United States of America,2014,111(11):4203-4208.
• [22]KE K,SUL O J,RAJASEKARAN M,et al.Micro RNA-183 increases osteoclastogenesis by repressing heme oxygenase-1[J].Bone,2015,81:237-246.
• [23]JIAO Y,ZHENG Z,DU X D,et al.Identification and characterization of micro RNAs in pearl oyster Pinctada martensii by Solexa deep sequencing[J].Marine Biotechnology,2014,16(1):54-62.
• [24]ZHENG Z,DU X D,XIONG X W,et al.Pm Runt regulated by Pm-mi R-183 participates in nacre formation possibly through promoting the expression of collagen VI-like and Nacrein in pearl oyster Pinctada martensii[J].PLo S One,2017,12(6):e0178561.
• [25]YUE F,ZHOU Z,WANG L L,et al.The essential roles of core binding factors Cf Runt and Cf CBFβin hemocyte production of scallop Chlamys farreri[J].Developmental&Comparative Immunology,2014,44(2):291-302.
• [26]DU X D,FAN G Y,JIAO Y,et al.The pearl oyster Pinctada fucata martensii genome and multi-omic analyses provide insights into biomineralization[J].Gigascience,2017,6(8):1-12.
• [27]ZUKER M.Mfold web server for nucleic acid folding and hybridization prediction[J].Nucleic Acids Research,2003,31(13):3406-3415.
• [28]KOZOMARA A,GRIFFITHS-JONES S.mi RBase:integrating micro RNA annotation and deep-sequencing data[J].Nucleic Acids Research,2011,39(Database issue):D152-D157.
• [29]郑哲.马氏珠母贝珍珠质形成micro RNA的筛选和功能研究[D].湛江:广东海洋大学,2013.
• [30]董冰冰.马氏珠母贝珍珠层基质蛋白基因的克隆与功能研究[D].湛江:广东海洋大学,2015.
• [31]雷倩楠.马氏珠母贝植核育珠移植耐受免疫相关基因与蛋白质鉴定及分析[D].湛江:广东海洋大学,2016.
• [32]鹿向东.mi R-146a靶向调控MIF基因对胶质瘤细胞增殖的影响[D].济南:山东大学,2016.
• [33]ESTELLER M.Non-coding RNAs in human disease[J].Nature Reviews Genetics,2011,12(12):861-874.
• [34]XU S B,WITMER P D,LUMAYAG S,et al.Micro RNA(mi RNA)transcriptome of mouse retina and identification of a sensory organ-specific mi RNA cluster[J].Journal of Biological Chemistry,2007,282(34):25053-25066.
• [35]DAVIS C,DUKES A,DREWRY M,et al.Micro RNA-183-5p increases with age in bone-derived extracellular vesicles,suppresses bone marrow stromal(stem)cell proliferation,and induces stem cell senescence[J].Tissue Engineering Part A,2017,23(21/22):1231-1240.
• [36]YAN F,LUO S J,JIAO Y,et al.Molecular characterization of the BMP7 gene and its potential role in shell formation in Pinctada martensii[J].International Journal of Molecular Sciences,2014,15(11):21215-21228.
• [37]DALUISKI A,ENGSTRAND T,BAHAMONDE M E,et al.Bone morphogenetic protein-3 is a negative regulator of bone density[J].Nature Genetics,2001,27(1):84-88.
• [38]MARIE B,JOUBERT C,BELLIARD C,et al.Characterization of MRNP34,a novel methionine-rich nacre protein from the pearl oysters[J].Amino Acids,2012,42(5):2009-2017.
• [39]TSUKAMOTO D,SARASHINA I,ENDO K.Structure and expression of an unusually acidic matrix protein of pearl oyster shells[J].Biochemical and Biophysical Research Communications,2004,320(4):1175-1180.