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http://dx.doi.org/10.5483/BMBRep.2015.48.10.098

Knockdown of microtubule actin crosslinking factor 1 inhibits cell proliferation in MC3T3-E1 osteoblastic cells  

Hu, Lifang (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Su, Peihong (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Li, Runzhi (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Yan, Kun (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Chen, Zhihao (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Shang, Peng (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Qian, Airong (Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University)
Publication Information
BMB Reports / v.48, no.10, 2015 , pp. 583-588 More about this Journal
Abstract
Microtubule actin crosslinking factor 1 (MACF1), a widely expressed cytoskeletal linker, plays important roles in various cells by regulating cytoskeleton dynamics. However, its role in osteoblastic cells is not well understood. Based on our previous findings that the association of MACF1 with F-actin and microtubules in osteoblast-like cells was altered under magnetic force conditions, here, by adopting a stable MACF1-knockdown MC3T3-E1 osteoblastic cell line, we found that MACF1 knockdown induced large cells with a binuclear/multinuclear structure. Further, immunofluorescence staining showed disorganization of F-actin and microtubules in MACF1-knockdown cells. Cell counting revealed significant decrease of cell proliferation and cell cycle analysis showed an S phase cell cycle arrest in MACF1-knockdown cells. Moreover and interestingly, MACF1 knockdown showed a potential effect on cellular MTT reduction activity and mitochondrial content, suggesting an impact on cellular metabolic activity. These results together indicate an important role of MACF1 in regulating osteoblastic cell morphology and function.
Keywords
Cell morphology; Cell proliferation; Cytoskeleton; MACF1; Osteoblastic cell;
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1 Louis KS and Siegel AC (2011) Cell viability analysis using trypan blue: manual and automated methods. Methods Mol Biol 740, 7-12   DOI
2 Wu X, Shen QT, Oristian DS et al (2011) Skin stem cells orchestrate directional migration by regulating microtubule-ACF7 connections through GSK3beta. Cell 144, 341-352   DOI
3 Sanchez-Soriano N, Travis M, Dajas-Bailador F, Goncalves-Pimentel C, Whitmarsh AJ and Prokop A (2009) Mouse ACF7 and drosophila short stop modulate filopodia formation and microtubule organisation during neuronal growth. J Cell Sci 122, 2534-2542   DOI
4 Antonellis PJ, Pollock LM, Chou SW et al (2014) ACF7 is a hair-bundle antecedent, positioned to integrate cuticular plate actin and somatic tubulin. J Neurosci 34, 305-312   DOI
5 Qian AR, Hu LF, Gao X et al (2009) Large gradient high magnetic field affects the association of MACF1 with actin and microtubule cytoskeleton. Bioelectromagnetics 30, 545-555   DOI
6 Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65, 55-63   DOI
7 Suzuki R, Matsuno S, Sakagami H, Okada Y and Shirataki Y (2014) Search of new cytotoxic crude materials against human oral squamous cell carcinoma using 1H NMRbased metabolomics. Anticancer Res 34, 4117-4120
8 Al-Rubeai M, Chalder S, Bird R and Emery AN (1991) Cell cycle, cell size and mitochondrial activity of hybridoma cells during batch cultivation. Cytotechnology 7, 179-186   DOI
9 Pagliacci MC, Spinozzi F, Migliorati G et al (1993) Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: a further pitfall in the use of the MTT assay for evaluating cell growth and survival. Eur J Cancer 29A, 1573-1577   DOI
10 Kitami T, Logan DJ, Negri J et al (2012) A chemical screen probing the relationship between mitochondrial content and cell size. PLoS One 7, e33755   DOI
11 Fuchs E and Karakesisoglou I (2001) Bridging cytoskeletal intersections. Genes Dev 15, 1-14   DOI
12 Kodama A, Karakesisoglou I, Wong E, Vaezi A and Fuchs E (2003) ACF7: an essential integrator of microtubule dynamics. Cell 115, 343-354   DOI
13 Leung CL, Green KJ and Liem RK (2002) Plakins: a family of versatile cytolinker proteins. Trends Cell Biol 12, 37-45   DOI
14 Leung CL, Sun D, Zheng M, Knowles DR and Liem RK (1999) Microtubule actin cross-linking factor (MACF): a hybrid of dystonin and dystrophin that can interact with the actin and microtubule cytoskeletons. J Cell Biol 147, 1275-1286   DOI
15 Sun D, Leung CL and Liem RK (2001) Characterization of the microtubule binding domain of microtubule actin crosslinking factor (MACF): identification of a novel group of microtubule associated proteins. J Cell Sci 114, 161-172
16 Karakesisoglou I, Yang Y and Fuchs E (2000) An epidermal plakin that integrates actin and microtubule networks at cellular junctions. J Cell Biol 149, 195-208   DOI
17 Bernier G, Pool M, Kilcup M, Alfoldi J, De Repentigny Y and Kothary R (2000) Acf7 (MACF) is an actin and microtubule linker protein whose expression predominates in neural, muscle, and lung development. Dev Dyn 219, 216-225   DOI
18 Ka M, Jung EM, Mueller U and Kim WY (2014) MACF1 regulates the migration of pyramidal neurons via microtubule dynamics and GSK-3 signaling. Dev Biol 395, 4-18   DOI
19 Wu X, Kodama A and Fuchs E (2008) ACF7 regulates cytoskeletal-focal adhesion dynamics and migration and has ATPase activity. Cell 135, 137-148   DOI
20 Qian AR, Gao X, Zhang W et al (2013) Large Gradient High Magnetic Fields Affect Osteoblast Ultrastructure and Function by Disrupting Collagen I or Fibronectin/alphabeta1 Integrin. PLoS One 8, e51036   DOI
21 Moulding DA, Blundell MP, Spiller DG et al (2007) Unregulated actin polymerization by WASp causes defects of mitosis and cytokinesis in X-linked neutropenia. J Exp Med 204, 2213-2224   DOI
22 Hossain MM, Hwang DY, Huang QQ, Sasaki Y and Jin JP (2003) Developmentally regulated expression of calponin isoforms and the effect of h2-calponin on cell proliferation. Am J Physiol Cell Physiol 284, C156-167   DOI
23 Zhu J, Beattie EC, Yang Y, Wang HJ, Seo JY and Yang LX (2005) Centrosome impairments and consequent cytokinesis defects are possible mechanisms of taxane drugs. Anticancer Res 25, 1919-1925
24 Menon MB and Gaestel M (2015) Sep(t)arate or not - how some cells take septin-independent routes through cytokinesis. J Cell Sci 128, 1877-1886   DOI
25 D'Avino PP, Giansanti MG and Petronczki M (2015) Cytokinesis in animal cells. Cold Spring Harb Perspect Biol 7, a015834   DOI
26 Livak KJ and Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT Method. Methods 25, 402-408   DOI
27 Qian A, Zhang W, Weng Y et al (2008) Gravitational environment produced by a superconducting magnet affects osteoblast morphology and functions. Acta Astronautica 63, 929-946   DOI
28 Mundra JJ, Terskiy A and Howells RD (2012) Naltrindole inhibits human multiple myeloma cell proliferation in vitro and in a murine xenograft model in vivo. J Pharmacol Exp Ther 342, 273-287   DOI