Effect of platelet-derived growth factor-BB on rat corpus cavernosum smooth muscle cell proliferation, migration and phenotypic modulation.
- Author:
Fengzhi CHEN
1
;
Shuhua HE
;
Haitao SHAN
;
Haibo ZHANG
;
Yanbing LIAN
;
Anyang WEI
Author Information
- Publication Type:Journal Article
- MeSH: Actins; metabolism; Animals; Cell Movement; drug effects; Cell Proliferation; drug effects; Cells, Cultured; Down-Regulation; Male; Myocytes, Smooth Muscle; cytology; drug effects; Myosin Heavy Chains; metabolism; Nuclear Proteins; metabolism; Penis; cytology; Phenotype; Proto-Oncogene Proteins c-sis; pharmacology; RNA, Messenger; Rats; Rats, Wistar; Trans-Activators; metabolism
- From: Journal of Southern Medical University 2015;35(7):971-976
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of platelet-derived growth factor-BB (PDGFBBB) on rat corpus cavernosum smooth muscle (CCSM) cell proliferation, migration and phenotypic modulation and explore the underlying mechanisms.
METHODSWistar rat CCSM cells were obtained through a modified tissue culture method and identified by immunofluorescence assay. The effect of PDGFBB on the proliferation of CCSM cells was investigated using a CCK-8 kit and the optimum PDGFBB concentration for cell treatment was determined. CCSM cells were treated with vehicle or PDGF-BB at the optimum concentration, and the cell migration was examined using scratch assay; the mRNA expression of the transcription factor myocardin and the contractile phenotype markers αSMA and SMMHC in CCSM cells were determined by qRT-PCR at 24 h and 48 h. The protein expression of myocardin in CCSM cells incubated with PDGFBB for 0, 24 and 48 h was examined by Western blotting.
RESULTIn CCSM cell culture, 96.5%and 96% of the cells were positive for αSMA and smoothelin, respectively. PDGFBB at different concentrations markedly promoted the proliferation of CCSM cells; the optimum PDGFBB concentration for enhancing cell proliferation was 12.5 ng/mL, which induced the migration of CCSM cells and significantly reduced the mRNA expressions of myocardin, αSMA and SMMHC (P<0.01). Exposure to PDGFBB decreased the protein expression of myocardin as the exposure time extended (within 48 h).
CONCLUSIONCCSM cells of a high purity can be obtained by the modified tissue culture method. PDGFBB can promote the proliferation and migration of CCSM cells and cause a phenotypic conversion from the contractile to the synthetic type possibly by down-regulating myocardin.