title: Molecular Mechanism of MicroRNA-200c Regulating Transforming Growth Factor-β (TGF-β)/SMAD Family Member 3 (SMAD3) Pathway by Targeting Zinc Finger E-Box Binding Homeobox 1 (ZEB1) in Hypospadias in Rats.
abstract: BACKGROUND The aim of this study was to explore effects of microRNA-200c regulating TGF-β/Smad3 pathway by targeting Zeb1 on the occurrence and development of hypospadias and to evaluate the relationship between microRNA-200c and occurrence of hypospadias. MATERIAL AND METHODS Pregnant rats with a gestational age of 12 days were allocated into 2 groups; one received gavage of DEHP-contained soybean oil (1 ml/day, 8 days; Group A) and the other had gavage of normal soybean oil (1 ml/day, 8 days; Group B). Baby rats with hypospadias from Group A were assigned to the model group (n=20) and healthy baby rats from Group B were assigned to the control group (n=20). Real-time quantitative polymerase chain reaction (qRT-PCR), immunohistochemistry and Western blot analysis were performed to detect microRNA-200c, Zeb1, TGF-β, and Smad3 mRNA and protein expressions in the model group (n=20) and the control group (n=20). The relationship between microRNA-200c and Zeb1 was detected using a dual-luciferase reporter gene experiment. After the in vitro intervention experiment in fetal rat penises, Western blot was used to detect the expression of Zeb1, TGF-β, and Smad3. RESULTS In the model group, microRNA-200c was expressed at a low level, and microRNA-200c expression in control group was 2.1 times higher than in the model group (P<0.05). When compared with the control group, mRNA expressions, protein expressions, and positive rates of Zeb1, TGF-β, and Smad3 were higher in the model group (all P<0.01). Luciferase gene report determined that Zeb1 is a target gene of microRNA-200c. The in vitro intervention experiment in fetal rat penises found that a high concentration of microRNA-200c inhibited hypospadias occurrence by suppressing the expression of Zeb1, TGF-β, and Smad3. CONCLUSIONS MicroRNA-200c was expressed in hypospadias penis tissues at low levels and was negatively correlated with Zeb1 expression. MicroRNA-200c up-regulated Zeb1 expression to regulate the TGF-β/Smad3 pathway, which led to the occurrence of hypospadias.
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Chong
authors: C
authors: Qian
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Xiangyang
authors: X
authors: Dang
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Xianglin
authors: X
authors: Wang
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Wei
authors: W
authors: Xu
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Guijian
authors: G
authors: Pang
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Yifeng
authors: Y
authors: Chen
authors: Department of Urology, The First People's Hospital of Yulin, Yulin, Guangxi, China (mainland).
authors: Chengbei
authors: C
authors: Liu
mesh_terms: D000818:Animals; D015536:Down-Regulation; D005801:Genes, Homeobox; D007021:Hypospadias / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D000071799:Zinc Finger E-box-Binding Homeobox 1 / Q000378:metabolism
chemical_list: C555600:MIRN200 microRNA, rat; D035683:MicroRNAs; D012333:RNA, Messenger; D051900:Smad3 Protein; C497787:Smad3 protein, rat; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D000071799:Zinc Finger E-box-Binding Homeobox 1