miR-195-5p Regulates the Phenotype Switch of CCSM Cells by Targeting Smad7

• Main Authors: Jing Zhang, MSc, Xingyuan Zhang, MS, Shengnan Cong, MS, Jingjing Zhang, MSc, Aixia Zhang, MD, Lianjun Pan, MD, Jiehua Ma, PhD
• Format: Article
• Language: English
• Published: Elsevier 2021-06-01
• Series: Sexual Medicine
• Subjects: Erectile Dysfunction (ED); miR-195-5p; Small Mother Against Decapentaplegic 7(Smad7); Corpus Cavernosum Smooth Muscle (CCSM); Phenotypic Transformation
• Online Access: http://www.sciencedirect.com/science/article/pii/S2050116121000295

ABSTRACT: Introduction: Phenotype switch refers to the process in which smooth muscle cells change from contractile type to synthetic type and acquire the ability of proliferation. Phenotypic transformation involves many changes of cell function, such as collagen deposition and fibrosis, which affect the normal erectile function of penis. Aim: To investigate the role of miR-195-5p in regulating the Phenotype switch of the corpus cavernosum smooth muscle (CCSM) cells. Methods: A small mother against decapentaplegic 7(Smad7) virus vector and a miR-195-5p mimics or an si-Smad7 viral vector and a miR-195-5p inhibitor were transfected into CCSM cells. The cells were obtained by primary culture of rat corpus cavernosum smooth muscle tissue. Real-time polymerase chain reaction (PCR) experiments, Western blotting, hematoxylin-eosin (HE) staining, transwell experiments, MTT assays, and flow cytometry were used to detect miR-195-5p, Smad7, phenotype switch markers of CCSM cells and related protein expression, as well as changes in cell morphology, migration, proliferation and apoptosis. Main Outcome Measure: To study the regulation of miR-195-5p in CCSM cells by overexpression and silencing strategies. Results: Overexpressed miR-195-5p promoted the transformation of CCSM cells from a contractile type to a synthetic type. Meanwhile, the migration ability and proliferation ability of CCSM cells increased, and the apoptosis rate decreased. The expression-silencing of miR-195-5p gave rise to the opposite effect. The results of the rescue experiment demonstrated that overexpressed Smad7 rescued the inhibitory of the switch of the CCSM cell phenotype from the contractile type to the synthesis type caused by overexpression of miR-195-5p alone. Moreover, the enhancement effect of the migration ability and proliferation ability of CCSM cells was also eliminated, and the apoptosis rate was increased. Silencing miR-195-5p and Smad7 at the same time resulted in the opposite effect. Conclusion: miR-195-5p may regulate the phenotype switch of CCSM cells by targeting Smad7.Zhang J, Zhang X, Zhang J, et al. miR-195-5p Regulates the Phenotype Switch of CCSM Cells by Targeting Smad7. Sex Med 2021;9:100349.