title: The Wnt11 Signaling Pathway in Potential Cellular EMT and Osteochondral Differentiation Progression in Nephrolithiasis Formation.
abstract: The molecular events leading to nephrolithiasis are extremely complex. Previous studies demonstrated that calcium and transforming growth factor-β1 (TGF-β1) may participate in the pathogenesis of stone formation, but the explicit mechanism has not been defined. Using a self-created genetic hypercalciuric stone-forming (GHS) rat model, we observed that the increased level of serous/uric TGF-β1 and elevated intracellular calcium in primary renal tubular epithelial cells (PRECs) was associated with nephrolithiasis progression in vivo. In the setting of high calcium plus high TGF-β1 in vitro, PRECs showed great potential epithelial to mesenchymal transition (EMT) progression and osteochondral differentiation properties, representing the multifarious increased mesenchymal and osteochondral phenotypes (Zeb1, Snail1, Col2A1, OPN, Sox9, Runx2) and decreased epithelial phenotypes (E-cadherin, CK19) bythe detection of mRNAs and corresponding proteins. Moreover, TGF-β-dependent Wnt11 knockdown and L-type Ca2+ channel blocker could greatly reverse EMT progression and osteochondral differentiation in PRECs. TGF-β1 alone could effectively promote EMT, but it had no effect on osteochondral differentiation in NRK cells (Rat kidney epithelial cell line). Stimulation with Ca2+ alone did not accelerate differentiation of NRK. Co-incubation of extracellular Ca2+ and TGF-β1 synergistically promotes EMT and osteochondral differentiation in NRK control cells. Our data supplied a novel view that the pathogenesis of calcium stone development may be associated with synergic effects of TGF-β1 and Ca2+, which promote EMT and osteochondral differentiation via Wnt11 and the L-type calcium channel.
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. hedeng2002@163.com.
authors.forename: Deng
authors.initials: D
authors.lastname: He
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. luyuchao@hust.edu.cn.
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. m201275806@hust.edu.cn.
authors.forename: Henglong
authors.initials: H
authors.lastname: Hu
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. medzjq@163.com.
authors.forename: Jiaqiao
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. qinbaolong@hust.edu.cn.
authors.forename: Baolong
authors.initials: B
authors.lastname: Qin
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. yfwangtjm@163.com.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. xingshuai529378226@126.com.
authors.forename: Shuai
authors.initials: S
authors.lastname: Xing
authors.affiliation: Department of Urology, the First Afﬁliated Hospital of Soochow University, Suzhou 215006, Jiangsu, China. linqix_827@163.com.
authors.forename: Qilin
authors.initials: Q
authors.lastname: Xi
authors.affiliation: Department of Urology, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. sgwangtjm@163.com.
authors.forename: Shaogang
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000473:pathology*; D020219:Chondrogenesis / Q000187:drug effects; D004195:Disease Models, Animal; D018450:Disease Progression; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D042541:Intracellular Space / Q000378:metabolism; D007684:Kidney Tubules / Q000473:pathology; D053040:Nephrolithiasis / Q000097:blood / Q000378:metabolism* / Q000473:pathology*; D009543:Nifedipine / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000097:blood / Q000652:urine; D051153:Wnt Proteins / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; D051153:Wnt Proteins; C502412:Wnt11 protein, rat; D009543:Nifedipine; D002118:Calcium
keywords: TGF-β1; Wnt11; calcium channel; nephrolithiasis