The function and mechanism of integrin signal pathway in the stretched rat BMSCs

Abstract

Abstract: PURPOSE: To detect the role and mechanism of integrin signal pathway acting on distraction osteogenesis（DO）. METHODS: A distraction model was established on the basis of an uniaxial static stretching device, and siRNA technology was used to silence the focal adhesion kinase (FAK) of bone marrow derived mesenchymal stem cells (BMCs), an important signal transduction molecule in integrin signaling pathway. After distraction, at 0, 6, 12 and 24 h, RNA and protein of important signal transduction molecules such as FAK, ILK and integrinβ1,were detected by RT-PCR and Western blot. And the cell proliferation,expression of the osteogenic genes Cbfa 1/Runx 2 and ALP were detected. SPSS 20.0 software package was used for one way analysis of variance. RESULTS: The expression of FAK specific siRNA lentiviral expression vector-bone differentiation factors Runx-2, ALP decreased compared to control group. And the expression of important signal molecules integrinβ1 and ILK in integrin pathway decreased dramatically. CONCLUSION: BMSCs bone differentiation is inhibited after FAK gene silence and the integrin signal pathway takes part in the conversion of cellar mechanical signals to biological chemical signals.

Key words: Distraction osteogenesis, BMSCs, Integrin signaling pathway, FAK

CLC Number:

Q257

JIANG Xi-liang, ZHU Xiao-wen, HU Jing, SONG Jian, YE Bing. The function and mechanism of integrin signal pathway in the stretched rat BMSCs[J]. China Journal of Oral and Maxillofacial Surgery, 2015, 13(3): 193-199.

References

[1] Amir LR, Everts V, Bronckers AL. Bone regeneration during distraction osteogenesis [J]. Odontology, 2009, 97(2): 63-75.
[2] Hibi H, Ueda M. Supraperiosteal transport distraction osteogenesis for reconstructing a segmental defect of the mandible[J]. J Oral Maxillofac Surg, 2011, 69(3): 742-746.
[3] Feng X, Tuo X, Chen F, et al. Ultrastructural cell response to tension stress during mandibular distraction osteogenesis [J]. Br J Oral Maxillofac Surg, 2008, 46(7): 527-532.
[4] 赵闯, 周昊, 朱晓文,等. Wnt/β-catenin信号通路在大鼠骨髓间充质干细胞牵张中的表达[J].中国口腔颌面外科杂志, 2013, 11(5): 354-358.
[5] Qi MC, Hu J, Zou SJ, et al. Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets -1 expression in stretched rat mesenchymal stem cells[J]. Int J Oral Maxillofac Surg, 2008, 37(5): 453-458.
[6] Ai-Aql ZS, Alagl AS, Graves DT, et al. Molecular mechanisms controlling bone formation during fracture healing and distraction osteogenesis [J]. J Dent Res, 2008, 87(2): 107-118.
[7] Wang Y, McNamara LM, Schaffler MB, et al. A model for the role of integrins in flow induced mechanotransduction in osteocytes [J]. Proc Natl Acad Sci USA, 2007, 104(40): 15941-15946.
[8] Spiteri C, Raizman I, Pilliar RM, et al. Matrix accumulation by articular chondrocytes during mechanical stimulation is influenced by integrin-mediated cell spreading[J]. J Biomed Mater Res A, 2010, 94(1): 122-129.
[9] Li J, Zhao Z, Wang J, et al. The role of extracellular matrix, integrins, and cytoskeleton in mechanotransduction of centrifugal loading [J]. Mol Cell Biochem, 2008, 309(1-2): 41-48.
[10] Puklin-Faucher E, Sheetz MP. The mechanical integrin cycle [J]. J Cell Sci, 2009, 15(122): 179-186.
[11] De Ugarte DA, Alfonso Z, Zuk PA, et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow[J]. Immunol Lett, 2003, 89(2-3): 267-270.
[12] Bouletreau PJ, Warren SM, Longaker MT. The molecular biology of distraction osteogenesis[J]. J Cranio-maxillofac Surg,2002,30(1): 1-11.
[13] Zhang M, Chen YJ, Ono T, et al. Crosstalk between integrin and G protein pathways involved in mechanotransduction in mandibular condylar chondrocytes under pressure [J]. Arch Biochem Biophys, 2008, 474(1): 102-108.
[14] Hynes Ro. Integrins: bidirectional, allosteric signaling machines[J]. Cell, 2002, 20(6): 673-687
[15] Kumar CC. Signaling by integrin receptors [J]. Oncogene, 1998, 17(11): 1365-1373.
[16] Salomon DS. Cell-cell and cell-extracellular matrix adhesion molecules communicate with growth factor receptors: an interactive signaling web [J].Cancer Invest, 2000, 18(6): 591-593.
[17] Nakagawa T, Yuan J. Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis [J]. J Cell Biol, 2000, 150(4): 887-894.
[18] Giancotti FG, Ruoslahti SE. Integrin signaling [J]. Science, 1999, 285(13): 1028-1032.
[19] Choi CK, Zareno J, Digman MA, et al. Cross-correlated fluctuation analysis reveals phosphorylation-regulated paxillin-FAK Complexes in nascent adhesions [J]. Biophys J, 2011,100(3): 583-592.
[20] Bol&#x0dbc0;&#x0dda9;s V, Gasent JM, L&#x0dbc0;&#x0dda9;pez-Tarruella S, et al. The dual kinase complex FAK-Src as a promising therapeutic target in cancer [J]. Onco Targets Ther, 2010, 3: 83-97.
[21] Zhao X, Guan JL. Focal adhesion kinase and its signaling pathways in cell migration and angiogenesis [J]. Adv Drug Deliv Rev, 2011, 63(8): 610-615.
[22] 杜菲, 黄常志. 整合素相关激酶(ILK)研究进展 [J]. 国外医学·分子生物学分册, 2001, 23(3): 142-145.
[23] 李继华, 胡静, 王志国, 等. 牵张成骨时整合素β1在细胞内外张应力传递中的作用 [J]. 口腔医学研究, 2005, 21(2): 127-129.
[24] Damsky CH, Ili D. Integrin signaling: it's where the action is [J]. Curr Opin Cell Biol, 2002, 14(5): 594-602.
[25] 张惠静, 卢晓, 蔡绍皙, 等. 整合素与肿瘤[J].肿瘤防治研究, 2002, 29(5): 425-427.
[26] 杨丽, 张荣华, 谢厚杰, 等. 建立大鼠骨髓间充质干细胞稳定分离培养体系与鉴定 [J].中国组织工程研究与临床康复, 2009, 13(6): 1064-1068.
[27] Novina CD, Sharp PA. The RNAi revolution [J].Nature,2004, 430(6996): 161-164.
[28] Agami R. RNAi and related mechanisms and their potential use for therapy [J].Curr Opin Chem Biol, 2002, 6(6): 829-834.
[29] Masiero M, Nardo G, Indraccolo S, et al. RNA interference: implications for cancer treatment[J]. Mol Aspects Med,2007,28(1): 143-166.
[30] 黎晓, 龚妍春.整合素及其信号转导通路的研究进展 [J]. 赣南医学院学报, 2007, 27(1): 153-154.
[31] 周建华, 许艳彬, 邱建忠, 等. 大鼠颌骨来源骨髓间充质干细胞中Cbfα1/p56亚型的表达 [J].中国组织工程研究, 2014, 18(23): 3622-3626.