{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,3]],"date-time":"2026-01-03T22:53:55Z","timestamp":1767480835350,"version":"build-2065373602"},"reference-count":33,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2015,7,17]],"date-time":"2015-07-17T00:00:00Z","timestamp":1437091200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science Foundation of P. R. China","award":["81170651","81270787"],"award-info":[{"award-number":["81170651","81270787"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The molecular events leading to nephrolithiasis are extremely complex.  Previous studies demonstrated that calcium and transforming growth factor-\u03b21 (TGF-\u03b21)  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-\u03b21 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-\u03b21 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-\u03b2-dependent Wnt11 knockdown and L-type Ca2+ channel blocker could greatly reverse EMT progression and osteochondral differentiation in PRECs.  TGF-\u03b21 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-\u03b21 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-\u03b21 and Ca2+, which promote EMT and osteochondral differentiation via Wnt11 and the L-type calcium channel.<\/jats:p>","DOI":"10.3390\/ijms160716313","type":"journal-article","created":{"date-parts":[[2015,7,17]],"date-time":"2015-07-17T11:18:48Z","timestamp":1437131928000},"page":"16313-16329","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["The Wnt11 Signaling Pathway in Potential Cellular EMT  and Osteochondral Differentiation Progression in Nephrolithiasis Formation"],"prefix":"10.3390","volume":"16","author":[{"given":"Deng","family":"He","sequence":"first","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuchao","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henglong","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6747-0654","authenticated-orcid":false,"given":"Jiaqiao","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baolong","family":"Qin","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yufeng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuai","family":"Xing","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qilin","family":"Xi","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Af\ufb01liated Hospital of Soochow University,  Suzhou 215006, Jiangsu, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaogang","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Urology, Tongji Hospital of Tongji Medical College of  Huazhong University of Science and Technology, Wuhan 430030, Hubei, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2015,7,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.1681\/ASN.2009060580","article-title":"Does idiopathic hypercalciuria trigger calcium-sensing receptor-mediated protection from urinary supersaturation?","volume":"20","author":"Worcester","year":"2009","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1681\/ASN.2004110991","article-title":"Regulation of renal calcium receptor gene expression by 1,25-dihydroxyvitamin D3 in genetic hypercalciuric stone-forming rats","volume":"16","author":"Yao","year":"2005","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1136\/jclinpath-2013-201879","article-title":"Male hypercalciuric stone formers with low renal calcium reabsorption","volume":"67","author":"Walker","year":"2014","journal-title":"J. Clin. Pathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"F853","DOI":"10.1152\/ajprenal.00124.2013","article-title":"Evidence for increased renal tubule and parathyroid gland sensitivity to serum calcium in human idiopathic hypercalciuria","volume":"305","author":"Worcester","year":"2013","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2589","DOI":"10.1210\/jc.2013-1301","article-title":"Hyperinsulinemia and urinary calcium excretion in calcium stone formers with idiopathic hypercalciuria","volume":"98","author":"Yoon","year":"2013","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/35036035","article-title":"The versatility and universality of calcium signalling","volume":"1","author":"Berridge","year":"2000","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"241","DOI":"10.5966\/sctm.2013-0079","article-title":"Human bone marrow-derived mesenchymal stem cells display enhanced clonogenicity but impaired differentiation with hypoxic preconditioning","volume":"3","author":"Boyette","year":"2014","journal-title":"Stem Cells Transl. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1002\/stem.1247","article-title":"Human bone marrow-derived mesenchymal stem cells suppress human glioma growth through inhibition of angiogenesis","volume":"31","author":"Ho","year":"2013","journal-title":"Stem Cells"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1390","DOI":"10.1016\/j.bbrc.2014.07.004","article-title":"High extracellular magnesium inhibits mineralized matrix deposition and modulates intracellular calcium signaling in human bone marrow-derived mesenchymal stem cells","volume":"450","author":"Zhang","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1472","DOI":"10.1096\/fj.06-7153com","article-title":"Physical manipulation of calcium oscillations facilitates osteodifferentiation of human mesenchymal stem cells","volume":"21","author":"Sun","year":"2007","journal-title":"FASEB J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1038\/nm1613","article-title":"Endothelial-to-mesenchymal transition contributes to cardiac fibrosis","volume":"13","author":"Zeisberg","year":"2007","journal-title":"Nat. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.bbrc.2014.05.125","article-title":"P44\/WDR77 restricts the sensitivity of proliferating cells to TGF \u03b2 signaling","volume":"450","author":"Yi","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"21290","DOI":"10.1074\/jbc.M112.357202","article-title":"Activation of Wnt11 by transforming growth factor-\u03b2 drives mesenchymal gene expression through non-canonical Wnt protein signaling in renal epithelial cells","volume":"287","author":"Zhang","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.urology.2013.11.004","article-title":"Does crystal deposition in genetic hypercalciuric rat kidney tissue share similarities with bone formation?","volume":"83","author":"Jia","year":"2014","journal-title":"Urology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.3892\/mmr.2014.2941","article-title":"Calcium ions promote primary renal epithelial cell differentiation into cells with bone-associated phenotypes via transforming growth factor-\u03b21-induced epithelial-mesenchymal transition in idiopathic hypercalciuria patients","volume":"11","author":"He","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1681\/ASN.2004100888","article-title":"Genetic hypercalciuria","volume":"16","author":"Moe","year":"2005","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1038\/ki.2008.425","article-title":"EMT and proteinuria as progression factors","volume":"75","author":"Strutz","year":"2009","journal-title":"Kidney Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1097\/01.ASN.0000106015.29070.E7","article-title":"Epithelial to mesenchymal transition in renal fibrogenesis: Pathologic significance, molecular mechanism, and therapeutic intervention","volume":"15","author":"Liu","year":"2004","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"681","DOI":"10.2169\/internalmedicine.47.0906","article-title":"Chronic kidney disease in Japan","volume":"47","author":"Iseki","year":"2008","journal-title":"Int. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1053\/j.ajkd.2009.08.008","article-title":"Risk factors for CKD in persons with kidney stones: A case-control study in Olmsted County, Minnesota","volume":"55","author":"Saucier","year":"2010","journal-title":"Am. J. Kidney Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3215","DOI":"10.1002\/dvdy.22459","article-title":"Canonical Wnt\/\u03b2-catenin signaling is required for maintenance but not activation of Pitx2 expression in neural crest during eye development","volume":"239","author":"Zacharias","year":"2010","journal-title":"Dev. Dyn."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1681\/ASN.2008060566","article-title":"Wnt\/\u03b2-catenin signaling promotes renal interstitial fibrosis","volume":"20","author":"He","year":"2009","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9298","DOI":"10.1158\/0008-5472.CAN-10-0226","article-title":"WNT11 expression is induced by estrogen-related receptor \u03b1 and \u03b2-catenin and acts in an autocrine manner to increase cancer cell migration","volume":"70","author":"Dwyer","year":"2010","journal-title":"Cancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1573","DOI":"10.1016\/j.cub.2009.07.062","article-title":"Wnt11\/5a complex formation caused by tyrosine sulfation increases canonical signaling activity","volume":"19","author":"Cha","year":"2009","journal-title":"Curr. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1007\/s12013-013-9639-z","article-title":"Secreted products of macrophages exposed to calcium oxalate crystals induce epithelial mesenchymal transition of renal tubular cells via RhoA-dependent TGF-\u03b21 pathway","volume":"67","author":"Kanlaya","year":"2013","journal-title":"Cell. Biochem. Biophys."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1007\/s00240-005-0509-z","article-title":"Elevated plasma concentrations of transforming growth factor-\u03b2 1 in patients with unilateral ureteral obstruction","volume":"33","author":"Vuruskan","year":"2005","journal-title":"Urol. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1083\/jcb.200303107","article-title":"Wnt-5\/pipetail functions in vertebrate axis formation as a negative regulator of Wnt\/\u03b2-catenin activity","volume":"162","author":"Westfall","year":"2003","journal-title":"J. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1007\/978-94-007-2888-2_46","article-title":"Mechanosensory calcium signaling","volume":"740","author":"Jones","year":"2012","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"6090","DOI":"10.1074\/jbc.275.9.6090","article-title":"Molecular and functional properties of the human \u03b11G subunit that forms T-type calcium channels","volume":"275","author":"Monteil","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"F997","DOI":"10.1152\/ajprenal.2000.279.6.F997","article-title":"The \u03b11G-subunit of a voltage-dependent Ca2+ channel is localized in rat distal nephron and collecting duct","volume":"279","author":"Andreasen","year":"2000","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1393","DOI":"10.1046\/j.1523-1755.2002.00267.x","article-title":"Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney","volume":"61","author":"Zhao","year":"2002","journal-title":"Kidney Int."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2661","DOI":"10.1681\/ASN.2006121410","article-title":"Reduced expression of insulin receptors in the kidneys of insulin-resistant rats","volume":"18","author":"Tiwari","year":"2007","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"F1055","DOI":"10.1152\/ajprenal.00108.2005","article-title":"Regulation of blood pressure, the epithelial sodium channel (ENaC), and other key renal sodium transporters by chronic insulin infusion in rats","volume":"290","author":"Song","year":"2006","journal-title":"Am. J. Physiol. Ren. Physiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/16\/7\/16313\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T20:49:19Z","timestamp":1760215759000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/16\/7\/16313"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,7,17]]},"references-count":33,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2015,7]]}},"alternative-id":["ijms160716313"],"URL":"https:\/\/doi.org\/10.3390\/ijms160716313","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2015,7,17]]}}}