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Journal of Southern Medical University ; (12): 971-976, 2015.
Article in Chinese | WPRIM | ID: wpr-355247

ABSTRACT

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Wistar 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.</p><p><b>RESULT</b>In 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).</p><p><b>CONCLUSION</b>CCSM 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.</p>


Subject(s)
Animals , Male , Rats , Actins , Metabolism , Cell Movement , Cell Proliferation , Cells, Cultured , Down-Regulation , Myocytes, Smooth Muscle , Cell Biology , Myosin Heavy Chains , Metabolism , Nuclear Proteins , Metabolism , Penis , Cell Biology , Phenotype , Proto-Oncogene Proteins c-sis , Pharmacology , RNA, Messenger , Rats, Wistar , Trans-Activators , Metabolism
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