The Changes of Cytoskeleton during Phenotypic Transition of Pulmonary Artery Smooth Muscle Cells Induced by PDGF-BB

WU Shan-shan, XIE Liang, LIU Han-min, LIU Bin

Abstract

To study the changes of cytoskeleton during the phenotypic transition of rat pulmonary artery smooth muscle cells (PASMCs) induced by platelet-derived growth factor (PDGF-BB), and to explore the mechanism involved in the process of phenotypic transition of PASMCs. Methods PASMCs of Sprague Dawley (SD) rats were cultured and identified by immunohistochemistry (IHC) method. The cells were randomly divided into control group and PDGF-BB treated group (10 ng/mL). RT-qPCR and Western blot were used to detect the mRNA and protein level of marker genes (α-SMA and SM22α) during the process of phenotypic transition of PASMCs. The changes of cytoskeleton were observed by fluorescent microscopy, cell proliferation was measured by CCK-8 method; and cell migration was observed by wound healing assay. Results Compared with control group, PDGF-BB down-regulated the mRNA and protein expression of α-SMA and SM22α. The fluorescence intensity of cytoskeletal protein was significantly reduced after the treatment of PDGF-BB. In addition, the structure of F-actin was disorganized with a burr-like appearance, and the structures of α-tubulin and β-tubulin were irregular with co-location appearance. PDGF-BB significantly enhanced the proliferation and migration of PASMCs. Conclusion PDGF-BB could induce a conformation change in cytoskeletal proteins for PASMCs phenotypic transition, and enhance the ability of proliferation and migration of PASMCs.

 

Keywords: Pulmonary artery smooth muscle cells, Phenotypic conversion, Cytoskeleton

 

Full Text:

PDF


References


BEAMISH JA, HE P. KOTTKEMARCHANT K, et al. Molecular regulation of contractile smooth muscle cell phenotype; implications for vascular tissue engineering. Tissue Eng Part В Rev,2010,16(5);467-491.

TAJSIC T, MORRELL NW. Smooth muscle cell hypertrophy, proliferation, migration and apoptosis in pulmonary hypertension. Compr Physiol. 2011, 1 ( 1 ); 295- 317.

SHIMODA LA. LAURIE SS. Vascular remodeling in pulmonary hypertension. J Mol Med (Berl) .2013.91(3);297- 309.

FREDRIKSSON L. LI H. ERIKSSON U. The PDGF family: four gene products form five dimeric isoforms. Cytokine Growth Factor Rev,2004,15(4): 197-204.

HASSOUN PM, MOUTHON L. BARBERA JA, et al. Inflammation, growth factors, and pulmonary vascular remodeling. J Am Coll Cardiol.2009,54(1 Suppl); 10-19.

SCHERMULY RT, DONY E, GHOFRANI HA, et al. Reversal of experimental pulmonary hypertension by PDGF inhibition. J Clin Invest, 2005,115( 10): 2811-2821.

CHEN S, LIU B, KONG D, et al. Atorvastatin calcium inhibits phenotypic modulation of PDGF-BB-induced VSMCs via down-regulation the Akt signaling pathway. PLoS One, 2015, 10 (4): e0122577 [ 2017-05-17]. https://doi. org/ 10. 1371/journal, pone. 0122577.

CHEN J, CUI X. QIAN Z, et al. Multi-omics analysis reveals regulators of the response to PDGF-BB treatment in pulmonary artery smooth muscle cells. BMC Genomics, 2016,17(1):781-796.

ZHAO Y. LV W, PIAO H. et al. Role of platelet-derived growth factor-BB (PDGF-BB) in human pulmonary artery smooth muscle cell proliferation. J Recept Signal Transduct Res,2014.34(4):254-260.

FAN Z, LI C, QIN C, et al. Role of the PI3K/AKT pathway in modulating cytoskeleton rearrangements and phenotype switching in rat pulmonary arterial vascular smooth muscle cells. DNA Cell Biol,2014,33( 1): 12-20.

YIN Y, WU X, YANG Z, et al. The potential efficacy of R8-modified paclitaxel-loaded liposomes on pulmonary arterial hypertension. Pharm Res,2013,30(8);2050-2062.

OWENS (Ж, KUMAR MS, WAMHOFF HR. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev,2004 ,84 (3); 767-801.

XIE C, GUO Y, ZHU T, et al. Yapl protein regulates vascular smooth muscle cell phenotypic switch by interaction with myocardin. J Biol Chem,2012,287( 18): 14598-14605.

HUANG J, PARMACEK MS. Modulation of smooth muscle cell phenotype: the other side of the story. Circ Res, 2012, 111(6):659-661.

WANG Z, WANG DZ, HOCKEMEYER D, et al. Myocardin and ternary complex factors compete for SRF to control smooth muscle gene expression. Nature, 2004, 428 (6979):185-189.

KAWAI-KOWASE K, OWENS GK. Multiple repressor pathways contribute to phenotypic switching of vascular smooth muscle cells. Am J Physiol Cell Physiol, 2007, 292 (1):C59-C69.

KAPLANALBUQUERQUE N, GARAT C, DESSEVA C, et al. Platelet-derived growth factor-bb-mediated activation of akt suppresses smooth muscle-specific gene expression through inhibition of mitogen-activated protein kinase and redistribution of serum response factor. J Biol Chem, 2003, 278(41):39830-39838.

FLETCHER DA, MULLINS RD. Cell mechanics and the cytoskeleton. Nature,2010,463(7280) :485-492.

TANG DD, GERLACH BD. The roles and regulation of the actin cytoskeleton, intermediate filaments and microtubules in smooth muscle cell migration. Respir Res,2017,18( 1) ;54- 66.

MACK CP, SOMLYO AV, HAUTMANN M. et al. Smooth muscle differentiation marker gene expression is regulated by RhoA-mediated act in polymerization. J Biol Chem.2001 • 276(1):341-347.

HELLSTRAND P. ALBINSSON S. Stretch-dependent growth and differentiation in vascular smooth muscle; role of the actin cytoskeleton. Can J Physiol Pharmacol, 2005, 83 (10):869-875.

HAN M, DONG LH, ZHENG B. et al. Smooth muscle 22 alpha maintains the differentiated phenotype of vascular smooth muscle cells by inducing filamentous actin bundling. Life Sci, 2009,84(13/14) s 394-401.

SON JE. LEE E, JUNG SK. et al. Anthocyanidins, novel FAK inhibitors, attenuate PDGF-BB-induced aortic smooth muscle cell migration and neointima formation. Cardiovasc Res,2014,101(3) ;503-512.

DONG L, WEN J, MIAO S, et al. Baicalin inhibits PDGF- BB-stimulated vascular smooth muscle cell proliferation through suppressing PDGFR^-ERK signaling and increase in p27 accumulation and prevents injury-induced neointimal hyperplasia. Cell Res,2010,20( 11), 1252-1262.


Refbacks

  • There are currently no refbacks.