miR-155通过下调BMP9/Smad信号通路抑制间充质干细胞C3H10T1/2成骨分化
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摘要
该文研究了在诱导小鼠间充质干细胞C3H10T1/2细胞成骨分化过程中miR-155的作用及其是否是通过调控BMP9/Smad(bonemorphogenetic protein 9/drosophila mothers against de-capentaplegic)信号通路发挥作用。在诱导C3H10T1/2细胞成骨分化过程中,采用实时定量PCR(Real-time quantitative PCR,q RT-PCR)检测miR-155水平的变化。转染miR-155模拟剂(miR-155)至C3H10T1/2细胞后,miR-155水平显著增高(P<0.001),而转染其抑制剂(anti-miR-155)至C3H10T1/2细胞后,miR-155水平显著降低(P<0.001)。转染后成骨诱导培养基诱导成骨7 d,碱性磷酸酶(alkaline phosphatase,ALP)活性及染色结果显示,miR-155能显著降低C3H10T1/2细胞成骨分化过程中的ALP活性(P<0.01)、减弱ALP染色,而anti-miR-155则能逆转其作用。成骨诱导14 d茜素红S染色结果显示,miR-155组钙盐结节较对照组少,下调miR-155的水平后,钙盐沉积结节增多。q RT-RCR检测结果显示,miR-155显著降低BMP9 m RNA水平(P<0.001),且miR-155组成骨基因Runx2和ALP表达均显著低于对照NC组(P<0.05、P<0.01)。Western blot检测BMP9、Runx2和p-Smad1/5/8蛋白质水平,结果显示,miR-155组蛋白质水平均显著降低(P<0.01、P<0.05、P<0.001)。该研究结果提示,miR-155对C3H10T1/2细胞成骨分化的抑制作用可能是通过抑制BMP9/Smad信号通路发挥作用的。
The aim of this study was to investigate the regulatory effect of miR-155 on the osteogenic differentiation of mesenchymal stem cells line C3H10T1/2 and its mechanisms. This study tested the miR-155 level during the process of osteogenic differentiation of C3H10T1/2 cells by Real-time quantitative PCR(q RT-PCR). Transfected with miR-155 mimic(miR-155) into C3H10T1/2 cells, the miR-155 level was significantly increased(P<0.001), however, when transfected with miR-155 inhibitor(anti-miR-155) into C3H10T1/2 cells, the miR-155 level was significantly decreased(P<0.001). After transfecting, induced osteogenic differentiation with osteoblast inducing conditional medium for 7 d, the results of Alkaline phosphatase(ALP) activity and ALP staining shown that miR-155 markedly decreased ALP activity(P<0.01) and attenuated ALP staining in the process of osteogenesis of C3H10T1/2 cells, however anti-miR-155 could reverse this effect. Induced osteogenic differentiation for 14 d, Alizarin red S staining shown that compared with negative control(NC) group, miR-155 notably reduced the deposition of calcium salt, and inhibition of miR-155 increased the deposition of calcium salt of C3H10T1/2 cells. It was also found that miR-155 significantly reduced the m RNA level of BMP9(P<0.001), and the m RNA levels of osteogenesis genes Runx2 and ALP decreased obviously(P<0.05, P<0.01). miR-155 decreased the protein levels of BMP9, Runx2 and p-Smad1/5/8 significantly(P<0.01, P<0.05, P<0.001). We concluded that the inhibitory effect of miR-155 on the osteogenic differentiation of C3H10T1/2 cells might through downregulated BMP9/Smad signal pathway.
The aim of this study was to investigate the regulatory effect of miR-155 on the osteogenic differentiation of mesenchymal stem cells line C3H10T1/2 and its mechanisms. This study tested the miR-155 level during the process of osteogenic differentiation of C3H10T1/2 cells by Real-time quantitative PCR(q RT-PCR). Transfected with miR-155 mimic(miR-155) into C3H10T1/2 cells, the miR-155 level was significantly increased(P<0.001), however, when transfected with miR-155 inhibitor(anti-miR-155) into C3H10T1/2 cells, the miR-155 level was significantly decreased(P<0.001). After transfecting, induced osteogenic differentiation with osteoblast inducing conditional medium for 7 d, the results of Alkaline phosphatase(ALP) activity and ALP staining shown that miR-155 markedly decreased ALP activity(P<0.01) and attenuated ALP staining in the process of osteogenesis of C3H10T1/2 cells, however anti-miR-155 could reverse this effect. Induced osteogenic differentiation for 14 d, Alizarin red S staining shown that compared with negative control(NC) group, miR-155 notably reduced the deposition of calcium salt, and inhibition of miR-155 increased the deposition of calcium salt of C3H10T1/2 cells. It was also found that miR-155 significantly reduced the m RNA level of BMP9(P<0.001), and the m RNA levels of osteogenesis genes Runx2 and ALP decreased obviously(P<0.05, P<0.01). miR-155 decreased the protein levels of BMP9, Runx2 and p-Smad1/5/8 significantly(P<0.01, P<0.05, P<0.001). We concluded that the inhibitory effect of miR-155 on the osteogenic differentiation of C3H10T1/2 cells might through downregulated BMP9/Smad signal pathway.
引文
1 Moffett HF,Novina CD.A small RNA makes a Bic difference.Genome Biol 2007;8(7):221.
2 Yanaihara N,Caplen N,Bowman E,Seike M,Kumamoto K,Yi M,et al.Unique micro RNA molecular profiles in lung cancer diagnosis and prognosis.Cancer Cell 2006;9(3):189-98.
3 Yang Y,Peng W,Tang T,Xia L,Wang XD,Duan BF,et al.Micro RNAs as promising biomarkers for tumor-staging:Evaluation of mi R21 mi R155 mi R29a and mi R92a in predicting tumor stage of rectal cancer.Asian Pac J Cancer Prev 2014;15(13):5175-80.
4 Wang Y,Keys DN,Au-Young JK,Chen C.Micro RNAs in embryonic stem cells.J Cell Physiol 2009;218(2):251-5.
5 Goff LA,Boucher S,Ricupero CL,Fenstermacher S,Swerdel M,Chase LG,et al.Differentiating human multipotent mesenchymal stromal cells regulate micro RNAs:Prediction of micro RNAregulation by PDGF during osteogenesis.Exp Hematol 2008;36(10):1354-69.
6 Sevli S,Uzumcu A,Solak M,Ittmann M,Ozen M.The function of micro RNAs,small but potent molecules,in human prostate cancer.Prostate Cancer Prostatic Dis 2010;13(3):208-17.
7 Eskildsen T,Taipaleenmaki H,Stenvang J,Abdallah BM,Ditzel N,Nossent AY,et al.Micro RNA-138 regulates osteogenic differentiation of human stromal(mesenchymal)stem cells in vivo.Proc Natl Acad Sci USA 2011;108(15):6139-44.
8 Li Z,Hassan MQ,Jafferji M,Aqeilan RI,Garzon R,Croce CM,et al.Biological functions of mi R-29b contribute to positive regulation of osteoblast differentiation.J Biol Chem 2009;284(23):15676-84.
9 Faraoni I,Antonetti FR,Cardone J,Bonmassar E.mi R-155 gene:A typical multifunctional micro RNA.Biochim Biophys Acta2009;1792(6):497-505.
10 Kluiver J,Poppema S,de Jong D,Blokzijl T,Harms G,Jacobs S,et al.BIC and mi R-155 are highly expressed in Hodgkin,primary mediastinal and diffuse large B cell lymphomas.J Pathol 2005;207(2):243-9.
11 Lawrie CH.Micro RNAs and lymphomagenesis:A functional review.Br J Haematol 2013;160(5):571-81.
12 Wang M,Tan LP,Dijkstra MK,van Lom K,Robertus JL,Harms G,et al.mi RNA analysis in B-cell chronic lymphocytic leukaemia:Proliferation centres characterized by low mi R-150and high BIC/mi R-155 expression.J Pathol 2008;215(1):13-20.
13 Jiang S,Zhang HW,Lu MH,He XH,Li Y,Gu H,et al.Micro RNA-155 functions as an Oncomi R in breast cancer by targeting the suppressor of cytokine signaling 1 gene.Cancer Res2010;70(8):3119-27.
14 Liu J,Mao Q,Liu Y,Hao X,Zhang S,Zhang J.Analysis of mi R-205 and mi R-155 expression in the blood of breast cancer patients.Chin J Cancer Res 2013;25(1):46-54.
15 Kong W,He L,Coppola M,Guo J,Esposito NN,Coppola D,et al.Micro RNA-155 regulates cell survival,growth,and chemosensitivity by targeting FOXO3a in breast cancer.J Biol Chem 2010;285(23):17869-79.
16 He B,Gao SQ,Huang LD,Huang YH,Zhang QY,Zhou MT,et al.Micro RNA-155 promotes the proliferation and invasion abilities of colon cancer cells by targeting quaking.Mol Med Rep2015;11(3):2355-9.
17 Dudda JC,Salaun B,Ji Y,Palmer DC,Monnot GC,Merck E,et al.Micro RNA-155 is required for effector CD8+T cell responses to virus infection and cancer.Immunity 2013;38(4):742-53.
18 Vigorito E,Perks KL,Abreu-Goodger C,Bunting S,Xiang Z,Kohlhaas S,et al.micro RNA-155 regulates the generation of immunoglobulin class-switched plasma cells.Immunity 2007;27(6):847-59.
19 Wang YZ,Feng XG,Shi QG,Hao YL,Yang Y,Zhang AM,et al.Silencing of mi R155 promotes the production of inflammatory mediators in Guillain-Barre syndrome in vitro.Inflammation2013;36(2):337-45.
20 O’Connell RM,Kahn D,Gibson WS,Round JL,Scholz RL,Chaudhuri AA,et al.Micro RNA-155 promotes autoimmune inflammation by enhancing inflammatory T cell development.Immunity 2010;33(4):607-19.
21 Du F,Yu F,Wang Y,Hui Y,Carnevale K,Fu M,et al.Micro RNA-155 deficiency results in decreased macrophage inflammation and attenuated atherogenesis in apolipoprotein E-deficient mice.Arterioscler Thromb Vasc Biol 2014;34(4):759-67.
22 Nazari-Jahantigh M,Wei Y,Noels H,Akhtar S,Zhou Z,Koenen RR,et al.Micro RNA-155 promotes atherosclerosis by repressing Bcl6 in macrophages.J Clin Invest 2012;122(11):4190-202.
23 Beederman M,Lamplot JD,Nan G,Wang J,Liu X,Yin L,et al.BMP signaling in mesenchymal stem cell differentiation and bone formation.J Biomed Sci Eng 2013;6(8A):32-52.
24 Brown MA,Zhao Q,Baker KA,Naik C,Chen C,Pukac L,et al.Crystal structure of BMP-9 and functional interactions with proregion and receptors.J Biol Chem 2005;280(26):25111-8.
25 Davis BN,Hilyard AC,Lagna G,Hata A.SMAD proteins control DROSHA-mediated micro RNA maturation.Nature 2008;454(7200):56-61.
26 Chen C,Tang Z,Song Q,Yang M,Shi Q,Weng Y.Downregulated micro RNA-23b promotes BMP9-mediated osteogenesis in C2C12 myoblast cells by targeting Runx2.Mol Med Rep2016;13(3):2492-8.
27 Song Q,Zhong L,Chen C,Tang Z,Liu H,Zhou Y,et al.mi R-21synergizes with BMP9 in osteogenic differentiation by activating the BMP9/Smad signaling pathway in murine multilineage cells.Int J Mol Med 2015;36(6):1497-506.
28 Zhang R,Weng Y,Li B,Jiang Y,Yan S,He F,et al.BMP9-induced osteogenic differentiation is partially inhibited by mi R-30a in the mesenchymal stem cell line C3H10T1/2.J Mol Histol2015;46(4/5):399-407.
29 Zhang W,Zhang L,Zhou Y,Ji X,Liu J,Liu D,et al.Synergistic effects of BMP9 and mi R-548d-5p on promoting osteogenic differentiation of mesenchymal stem cells.Biomed Res Int 2015;2015:309747.
30 Huang J,Zhao L,Xing L,Chen D.Micro RNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation.Stem cells 2010;28(2):357-64.
31 Inose H,Ochi H,Kimura A,Fujita K,Xu R,Sato S,et al.Amicro RNA regulatory mechanism of osteoblast differentiation.Proc Natl Acad Sci USA 2009;106(49):20794-9.
32 Itoh T,Nozawa Y,Akao Y.Micro RNA-141 and-200a are involved in bone morphogenetic protein-2-induced mouse preosteoblast differentiation by targeting distal-less homeobox 5.JBiol Chem 2009;284(29):19272-9.
33 Kang H,Louie J,Weisman A,Sheu-Gruttadauria J,DavisDusenbery BN,Lagna G,et al.Inhibition of micro RNA-302(mi R-302)by bone morphogenetic protein 4(BMP4)facilitates the BMP signaling pathway.J Biol Chem 2012;287(46):38656-64.
34 Kureel J,Dixit M,Tyagi AM,Mansoori MN,Srivastava K,Raghuvanshi A,et al.mi R-542-3p suppresses osteoblast cell proliferation and differentiation,targets BMP-7 signaling and inhibits bone formation.Cell Death Dis 2014;5:e1050.
35 Li Z,Hassan MQ,Volinia S,van Wijnen AJ,Stein JL,Croce CM,et al.A micro RNA signature for a BMP2-induced osteoblast lineage commitment program.Proc Natl Acad Sci USA 2008;105(37):13906-11.
36 Mizuno Y,Yagi K,Tokuzawa Y,Kanesaki-Yatsuka Y,Suda T,Katagiri T,et al.mi R-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation.Biochem Biophys Res Commun 2008;368(2):267-72.
37杨晓庆,苏健.mi R-155在骨髓间充质干细胞成骨分化中的调控作用.中国医药学导报(Yang Xiaoqing,Su Jian.Regulation effect of mi R-155 in the osteogenic differentiation of bone marrow mesenchymal stem cells.China Medical Herald)2015;12(33):38-6.
38 Shui W,Zhang W,Yin L,Nan G,Liao Z,Zhang H,et al.Characterization of scaffold carriers for BMP9-transduced osteoblastic progenitor cells in bone regeneration.J Biomed Mater Res A 2014;102(10):3429-38.
39 Li JZ,Hankins GR,Kao C,Li H,Kammauff J,Helm GA.Osteogenesis in rats induced by a novel recombinant helperdependent bone morphogenetic protein-9(BMP-9)adenovirus.JGene Med 2003;5(9):748-56.
40 Zhao Y,Song T,Wang W,Wang J,He J,Wu N,et al.P38 and ERK1/2 MAPKs act in opposition to regulate BMP9-induced osteogenic differentiation of mesenchymal progenitor cells.PLo SOne 2012;7(8):e43383.
41 Zhao YF,Xu J,Wang WJ,Wang J,He JW,Li L,et al.Activation of JNKs is essential for BMP9-induced osteogenic differentiation of mesenchymal stem cells.BMB Rep 2013;46(8):422-7.
42 Yin Q,Wang X,Fewell C,Cameron J,Zhu H,Baddoo M,et al.Micro RNA mi R-155 inhibits bone morphogenetic protein(BMP)signaling and BMP-mediated Epstein-Barr virus reactivation.JVirol 2010;84(13):6318-27.
43 Cargnello M,Roux PP.Activation and function of the MAPKs and their substrates,the MAPK-activated protein kinases.Microbiol Mol Biol Rev 2011;75(1):50-83.
44 Tang N,Song WX,Luo J,Luo X,Chen J,Sharff KA,et al.BMP-9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signalling.J Cell Mol Med 2009;13(8B):2448-64.
2 Yanaihara N,Caplen N,Bowman E,Seike M,Kumamoto K,Yi M,et al.Unique micro RNA molecular profiles in lung cancer diagnosis and prognosis.Cancer Cell 2006;9(3):189-98.
3 Yang Y,Peng W,Tang T,Xia L,Wang XD,Duan BF,et al.Micro RNAs as promising biomarkers for tumor-staging:Evaluation of mi R21 mi R155 mi R29a and mi R92a in predicting tumor stage of rectal cancer.Asian Pac J Cancer Prev 2014;15(13):5175-80.
4 Wang Y,Keys DN,Au-Young JK,Chen C.Micro RNAs in embryonic stem cells.J Cell Physiol 2009;218(2):251-5.
5 Goff LA,Boucher S,Ricupero CL,Fenstermacher S,Swerdel M,Chase LG,et al.Differentiating human multipotent mesenchymal stromal cells regulate micro RNAs:Prediction of micro RNAregulation by PDGF during osteogenesis.Exp Hematol 2008;36(10):1354-69.
6 Sevli S,Uzumcu A,Solak M,Ittmann M,Ozen M.The function of micro RNAs,small but potent molecules,in human prostate cancer.Prostate Cancer Prostatic Dis 2010;13(3):208-17.
7 Eskildsen T,Taipaleenmaki H,Stenvang J,Abdallah BM,Ditzel N,Nossent AY,et al.Micro RNA-138 regulates osteogenic differentiation of human stromal(mesenchymal)stem cells in vivo.Proc Natl Acad Sci USA 2011;108(15):6139-44.
8 Li Z,Hassan MQ,Jafferji M,Aqeilan RI,Garzon R,Croce CM,et al.Biological functions of mi R-29b contribute to positive regulation of osteoblast differentiation.J Biol Chem 2009;284(23):15676-84.
9 Faraoni I,Antonetti FR,Cardone J,Bonmassar E.mi R-155 gene:A typical multifunctional micro RNA.Biochim Biophys Acta2009;1792(6):497-505.
10 Kluiver J,Poppema S,de Jong D,Blokzijl T,Harms G,Jacobs S,et al.BIC and mi R-155 are highly expressed in Hodgkin,primary mediastinal and diffuse large B cell lymphomas.J Pathol 2005;207(2):243-9.
11 Lawrie CH.Micro RNAs and lymphomagenesis:A functional review.Br J Haematol 2013;160(5):571-81.
12 Wang M,Tan LP,Dijkstra MK,van Lom K,Robertus JL,Harms G,et al.mi RNA analysis in B-cell chronic lymphocytic leukaemia:Proliferation centres characterized by low mi R-150and high BIC/mi R-155 expression.J Pathol 2008;215(1):13-20.
13 Jiang S,Zhang HW,Lu MH,He XH,Li Y,Gu H,et al.Micro RNA-155 functions as an Oncomi R in breast cancer by targeting the suppressor of cytokine signaling 1 gene.Cancer Res2010;70(8):3119-27.
14 Liu J,Mao Q,Liu Y,Hao X,Zhang S,Zhang J.Analysis of mi R-205 and mi R-155 expression in the blood of breast cancer patients.Chin J Cancer Res 2013;25(1):46-54.
15 Kong W,He L,Coppola M,Guo J,Esposito NN,Coppola D,et al.Micro RNA-155 regulates cell survival,growth,and chemosensitivity by targeting FOXO3a in breast cancer.J Biol Chem 2010;285(23):17869-79.
16 He B,Gao SQ,Huang LD,Huang YH,Zhang QY,Zhou MT,et al.Micro RNA-155 promotes the proliferation and invasion abilities of colon cancer cells by targeting quaking.Mol Med Rep2015;11(3):2355-9.
17 Dudda JC,Salaun B,Ji Y,Palmer DC,Monnot GC,Merck E,et al.Micro RNA-155 is required for effector CD8+T cell responses to virus infection and cancer.Immunity 2013;38(4):742-53.
18 Vigorito E,Perks KL,Abreu-Goodger C,Bunting S,Xiang Z,Kohlhaas S,et al.micro RNA-155 regulates the generation of immunoglobulin class-switched plasma cells.Immunity 2007;27(6):847-59.
19 Wang YZ,Feng XG,Shi QG,Hao YL,Yang Y,Zhang AM,et al.Silencing of mi R155 promotes the production of inflammatory mediators in Guillain-Barre syndrome in vitro.Inflammation2013;36(2):337-45.
20 O’Connell RM,Kahn D,Gibson WS,Round JL,Scholz RL,Chaudhuri AA,et al.Micro RNA-155 promotes autoimmune inflammation by enhancing inflammatory T cell development.Immunity 2010;33(4):607-19.
21 Du F,Yu F,Wang Y,Hui Y,Carnevale K,Fu M,et al.Micro RNA-155 deficiency results in decreased macrophage inflammation and attenuated atherogenesis in apolipoprotein E-deficient mice.Arterioscler Thromb Vasc Biol 2014;34(4):759-67.
22 Nazari-Jahantigh M,Wei Y,Noels H,Akhtar S,Zhou Z,Koenen RR,et al.Micro RNA-155 promotes atherosclerosis by repressing Bcl6 in macrophages.J Clin Invest 2012;122(11):4190-202.
23 Beederman M,Lamplot JD,Nan G,Wang J,Liu X,Yin L,et al.BMP signaling in mesenchymal stem cell differentiation and bone formation.J Biomed Sci Eng 2013;6(8A):32-52.
24 Brown MA,Zhao Q,Baker KA,Naik C,Chen C,Pukac L,et al.Crystal structure of BMP-9 and functional interactions with proregion and receptors.J Biol Chem 2005;280(26):25111-8.
25 Davis BN,Hilyard AC,Lagna G,Hata A.SMAD proteins control DROSHA-mediated micro RNA maturation.Nature 2008;454(7200):56-61.
26 Chen C,Tang Z,Song Q,Yang M,Shi Q,Weng Y.Downregulated micro RNA-23b promotes BMP9-mediated osteogenesis in C2C12 myoblast cells by targeting Runx2.Mol Med Rep2016;13(3):2492-8.
27 Song Q,Zhong L,Chen C,Tang Z,Liu H,Zhou Y,et al.mi R-21synergizes with BMP9 in osteogenic differentiation by activating the BMP9/Smad signaling pathway in murine multilineage cells.Int J Mol Med 2015;36(6):1497-506.
28 Zhang R,Weng Y,Li B,Jiang Y,Yan S,He F,et al.BMP9-induced osteogenic differentiation is partially inhibited by mi R-30a in the mesenchymal stem cell line C3H10T1/2.J Mol Histol2015;46(4/5):399-407.
29 Zhang W,Zhang L,Zhou Y,Ji X,Liu J,Liu D,et al.Synergistic effects of BMP9 and mi R-548d-5p on promoting osteogenic differentiation of mesenchymal stem cells.Biomed Res Int 2015;2015:309747.
30 Huang J,Zhao L,Xing L,Chen D.Micro RNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation.Stem cells 2010;28(2):357-64.
31 Inose H,Ochi H,Kimura A,Fujita K,Xu R,Sato S,et al.Amicro RNA regulatory mechanism of osteoblast differentiation.Proc Natl Acad Sci USA 2009;106(49):20794-9.
32 Itoh T,Nozawa Y,Akao Y.Micro RNA-141 and-200a are involved in bone morphogenetic protein-2-induced mouse preosteoblast differentiation by targeting distal-less homeobox 5.JBiol Chem 2009;284(29):19272-9.
33 Kang H,Louie J,Weisman A,Sheu-Gruttadauria J,DavisDusenbery BN,Lagna G,et al.Inhibition of micro RNA-302(mi R-302)by bone morphogenetic protein 4(BMP4)facilitates the BMP signaling pathway.J Biol Chem 2012;287(46):38656-64.
34 Kureel J,Dixit M,Tyagi AM,Mansoori MN,Srivastava K,Raghuvanshi A,et al.mi R-542-3p suppresses osteoblast cell proliferation and differentiation,targets BMP-7 signaling and inhibits bone formation.Cell Death Dis 2014;5:e1050.
35 Li Z,Hassan MQ,Volinia S,van Wijnen AJ,Stein JL,Croce CM,et al.A micro RNA signature for a BMP2-induced osteoblast lineage commitment program.Proc Natl Acad Sci USA 2008;105(37):13906-11.
36 Mizuno Y,Yagi K,Tokuzawa Y,Kanesaki-Yatsuka Y,Suda T,Katagiri T,et al.mi R-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation.Biochem Biophys Res Commun 2008;368(2):267-72.
37杨晓庆,苏健.mi R-155在骨髓间充质干细胞成骨分化中的调控作用.中国医药学导报(Yang Xiaoqing,Su Jian.Regulation effect of mi R-155 in the osteogenic differentiation of bone marrow mesenchymal stem cells.China Medical Herald)2015;12(33):38-6.
38 Shui W,Zhang W,Yin L,Nan G,Liao Z,Zhang H,et al.Characterization of scaffold carriers for BMP9-transduced osteoblastic progenitor cells in bone regeneration.J Biomed Mater Res A 2014;102(10):3429-38.
39 Li JZ,Hankins GR,Kao C,Li H,Kammauff J,Helm GA.Osteogenesis in rats induced by a novel recombinant helperdependent bone morphogenetic protein-9(BMP-9)adenovirus.JGene Med 2003;5(9):748-56.
40 Zhao Y,Song T,Wang W,Wang J,He J,Wu N,et al.P38 and ERK1/2 MAPKs act in opposition to regulate BMP9-induced osteogenic differentiation of mesenchymal progenitor cells.PLo SOne 2012;7(8):e43383.
41 Zhao YF,Xu J,Wang WJ,Wang J,He JW,Li L,et al.Activation of JNKs is essential for BMP9-induced osteogenic differentiation of mesenchymal stem cells.BMB Rep 2013;46(8):422-7.
42 Yin Q,Wang X,Fewell C,Cameron J,Zhu H,Baddoo M,et al.Micro RNA mi R-155 inhibits bone morphogenetic protein(BMP)signaling and BMP-mediated Epstein-Barr virus reactivation.JVirol 2010;84(13):6318-27.
43 Cargnello M,Roux PP.Activation and function of the MAPKs and their substrates,the MAPK-activated protein kinases.Microbiol Mol Biol Rev 2011;75(1):50-83.
44 Tang N,Song WX,Luo J,Luo X,Chen J,Sharff KA,et al.BMP-9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signalling.J Cell Mol Med 2009;13(8B):2448-64.