title: Erbin interacts with Sema4C and inhibits Sema4C-induced epithelial-mesenchymal transition in HK2 cells.
abstract: Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Qiao-Dan
authors.initials: QD
authors.lastname: Zhou
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ning
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zeng
authors.affiliation: Department of Hepatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Kou
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Chu-Ou
authors.initials: CO
authors.lastname: Xu
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Guang-Chang
authors.initials: GC
authors.lastname: Pei
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Min
authors.initials: M
authors.lastname: Han
authors.affiliation: Department of Nephrology, Huazhong University of Science and Technology, Wuhan, 430030, China. xugang@tjh.tjmu.edu.cn.
authors.forename: Gang
authors.initials: G
authors.lastname: Xu
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002460:Cell Line; D058750:Epithelial-Mesenchymal Transition; D006801:Humans; D047468:Immunoprecipitation; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011485:Protein Binding; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D039961:Semaphorins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D014746:Vimentin / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D015820:Cadherins; C413364:ERBIN protein, human; C000592635:Sema4c protein, human; D039961:Semaphorins; D053773:Transforming Growth Factor beta1; D014746:Vimentin