Zhang Jing, Zhang Xingyuan, Cong Shengnan, Zhang Jingjing, Zhang Aixia, Pan Lianjun, Ma Jiehua
Jiangsu Health Vocational College, Nanjing, China.
Affiliated Obstetrics and Gynecology Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Center, Nanjing, China.
Sex Med. 2021 Jun;9(3):100349. doi: 10.1016/j.esxm.2021.100349. Epub 2021 Jun 1.
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.
To investigate the role of miR-195-5p in regulating the Phenotype switch of the corpus cavernosum smooth muscle (CCSM) cells.
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.
To study the regulation of miR-195-5p in CCSM cells by overexpression and silencing strategies.
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.
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.
表型转换是指平滑肌细胞从收缩型转变为合成型并获得增殖能力的过程。表型转变涉及细胞功能的许多变化,如胶原蛋白沉积和纤维化,这会影响阴茎的正常勃起功能。
探讨miR-195-5p在调节海绵体平滑肌(CCSM)细胞表型转换中的作用。
将小发夹RNA 7(Smad7)病毒载体与miR-195-5p模拟物或si-Smad7病毒载体与miR-195-5p抑制剂转染到CCSM细胞中。细胞通过大鼠海绵体平滑肌组织的原代培养获得。采用实时聚合酶链反应(PCR)实验、蛋白质印迹法、苏木精-伊红(HE)染色、Transwell实验、MTT法和流式细胞术检测miR-195-5p、Smad7、CCSM细胞的表型转换标志物及相关蛋白表达,以及细胞形态、迁移、增殖和凋亡的变化。
通过过表达和沉默策略研究miR-195-5p对CCSM细胞的调控作用。
过表达miR-195-5p促进CCSM细胞从收缩型向合成型转变。同时,CCSM细胞的迁移能力和增殖能力增强,凋亡率降低。miR-195-5p表达沉默则产生相反的效果。挽救实验结果表明,过表达Smad7可挽救由单独过表达miR-195-5p引起的CCSM细胞表型从收缩型向合成型转换的抑制作用。此外,CCSM细胞迁移能力和增殖能力的增强作用也被消除,凋亡率升高。同时沉默miR-195-5p和Smad7则产生相反的效果。
miR-195-5p可能通过靶向Smad7调节CCSM细胞的表型转换。张J,张X,张J等。miR-195-5p通过靶向Smad7调节CCSM细胞的表型转换。性医学2021;9:100349。