Lefty A protein inhibits TGF-β1-mediated apoptosis in human renal tubular epithelial cells

Zheng,Ren-Ping; Bai,Tao; Zhou,Xiao-Guang; Xu,Chang-Gen; Wang,Wei; Xu,Ming-Wei; Zhang,Jie

doi:10.3892/mmr.2013.1556

Lefty A protein inhibits TGF-β1-mediated apoptosis in human renal tubular epithelial cells

Authors:
- Ren-Ping Zheng
- Tao Bai
- Xiao-Guang Zhou
- Chang-Gen Xu
- Wei Wang
- Ming-Wei Xu
- Jie Zhang
View Affiliations

Affiliations: Department of Urology, Renmin Hospital of Wuhan University, Wuhan 430060, P.R. China
Published online on: June 26, 2013 https://doi.org/10.3892/mmr.2013.1556
Pages: 621-625

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

This study aimed to examine the effects of Lefty A protein on transforming growth factor-β1 (TGF-β1)-mediated apoptosis in human renal tubular epithelial cells (HK-2). HK-2 cells were transfected with the human Lefty gene to induce the secretion of endogenous Lefty A protein. Following exposure of the HK-2 cells to recombinant human TGF-β1 (10 ng/ml), p-Smad2/3 protein levels were examined by western blot analysis, and cellular apoptosis was detected by flow cytometry 6, 12, 24 and 48 h following TGF-β1 treatment. Coculture of renal tubular epithelial cells with TGF-β1 resulted in a significant increase in p-Smad2/3 protein levels and the rate of cell apoptosis, which were attenuated by liposome-mediated transfection with the Lefty gene. Lefty A protein was able to inhibit the TGF-β1/Smad signaling pathway and markedly attenuate TGF-β1-mediated apoptosis in human renal tubular epithelial cells. Taken together, these results indicated that the TGF-β1/Smad signaling pathway most likely mediates apoptosis in renal tubular epithelial cells. In addition, Lefty A protein is capable of inhibiting the TGF-β1/Smad pathway to reduce TGF-β1/Smad-mediated apoptosis in renal tubular epithelial cells. This study may provide novel insights into the prevention and treatment of urinary tract obstruction disease using Lefty A protein.

View Figures

View References

1	Padanilam BJ: Cell death induced by acute renal injury: a perspective on the contributions of apoptosis and necrosis. Am J Physiol Renal Physiol. 284:F608–F627. 2003. View Article : Google Scholar : PubMed/NCBI
2	Carson DA and Ribeiro JM: Apoptosis and disease. Lancet. 341:1251–1254. 1993. View Article : Google Scholar : PubMed/NCBI
3	Sánchez-Capelo A: Dual role for TGF-beta1 in apoptosis. Cytokine Growth Factor Rev. 16:15–34. 2005.
4	Rana A, Sathyanarayana P and Lieberthal W: Role of apoptosis of renal tubular cells in acute renal failure: therapeutic implications. Apoptosis. 6:83–102. 2001. View Article : Google Scholar : PubMed/NCBI
5	Havasi A and Borkan SC: Apoptosis and acute kidney injury. Kidney Int. 80:29–40. 2011. View Article : Google Scholar
6	Razzaque MS, Ahsan N and Taguchi T: Role of apoptosis in fibrogenesis. Nephron. 90:365–372. 2002. View Article : Google Scholar : PubMed/NCBI
7	Zeisberg M, Strutz F and Müller GA: Renal fibrosis: an update. Curr Opin Nephrol Hypertens. 10:315–320. 2001. View Article : Google Scholar
8	Böttinger EP and Bitzer M: TGF-beta signaling in renal disease. J Am Soc Nephrol. 13:2600–2610. 2002.
9	Kopp JB: TGF-beta signaling and the renal tubular epithelial cell: too much, too little, and just right. J Am Soc Nephrol. 21:1241–1243. 2010. View Article : Google Scholar : PubMed/NCBI
10	Meno C, Ito Y, Saijoh Y, et al: Two closely-related left-right asymmetrically expressed genes, lefty-1 and lefty-2: their distinct expression domains, chromosomal linkage and direct neuralizing activity in Xenopus embryos. Genes Cells. 2:513–524. 1997. View Article : Google Scholar
11	Meno C, Saijoh Y, Fujii H, et al: Left-right asymmetric expression of the TGF beta-family member lefty in mouse embryos. Nature. 381:151–155. 1996. View Article : Google Scholar : PubMed/NCBI
12	Kothapalli R, Buyuksal I, Wu SQ, Chegini N and Tabibzadeh S: Detection of ebaf, a novel human gene of the transforming growth factor beta superfamily association of gene expression with endometrial bleeding. J Clin Invest. 99:2342–2350. 1997. View Article : Google Scholar : PubMed/NCBI
13	Kosaki K, Bassi MT, Kosaki R, et al: Characterization and mutation analysis of human LEFTY A and LEFTY B, homologues of murine genes implicated in left-right axis development. Am J Hum Genet. 64:712–721. 1999. View Article : Google Scholar : PubMed/NCBI
14	Hamada H, Meno C, Watanabe D and Saijoh Y: Establishment of vertebrate left-right asymmetry. Nat Rev Genet. 3:103–113. 2002. View Article : Google Scholar : PubMed/NCBI
15	Dvash T, Mayshar Y, Darr H, et al: Temporal gene expression during differentiation of human embryonic stem cells and embryoid bodies. Hum Reprod. 19:2875–2883. 2004. View Article : Google Scholar : PubMed/NCBI
16	Marjoram L and Wright C: Rapid differential transport of Nodal and Lefty on sulfated proteoglycan-rich extracellular matrix regulates left-right asymmetry in Xenopus. Development. 138:475–485. 2011. View Article : Google Scholar : PubMed/NCBI
17	Ulloa L and Tabibzadeh S: Lefty inhibits receptor-regulated Smad phosphorylation induced by the activated transforming growth factor-beta receptor. J Biol Chem. 276:21397–21404. 2001. View Article : Google Scholar : PubMed/NCBI
18	Mason JM, Xu HP, Rao SK, et al: Lefty contributes to the remodeling of extracellular matrix by inhibition of connective tissue growth factor and collagen mRNA expression and increased proteolytic activity in a fibrosarcoma model. J Biol Chem. 277:407–415. 2002. View Article : Google Scholar
19	Miyajima A, Chen J, Lawrence C, et al: Antibody to transforming growth factor-beta ameliorates tubular apoptosis in unilateral ureteral obstruction. Kidney Int. 58:2301–2313. 2000. View Article : Google Scholar : PubMed/NCBI
20	Dai C, Yang J and Liu Y: Transforming growth factor-beta1 potentiates renal tubular epithelial cell death by a mechanism independent of Smad signaling. J Biol Chem. 278:12537–12545. 2003. View Article : Google Scholar : PubMed/NCBI
21	Li Y, Ge Y, Liu FY, et al: Norcantharidin, a protective therapeutic agent in renal tubulointerstitial fibrosis. Mol Cell Biochem. 361:79–83. 2012. View Article : Google Scholar : PubMed/NCBI
22	Wang X, Sun W, Zhang C, et al: TGF-beta1 inhibits the growth and metastasis of tongue squamous carcinoma cells through Smad4. Gene. 485:160–166. 2011. View Article : Google Scholar : PubMed/NCBI
23	Li J, Deane JA, Campanale NV, Bertram JF and Ricardo SD: Blockade of p38 mitogen-activated protein kinase and TGF-beta1/Smad signaling pathways rescues bone marrow-derived peritubular capillary endothelial cells in adriamycin-induced nephrosis. J Am Soc Nephrol. 17:2799–2811. 2006. View Article : Google Scholar
24	Inazaki K, Kanamaru Y, Kojima Y, et al: Smad3 deficiency attenuates renal fibrosis, inflammation, and apoptosis after unilateral ureteral obstruction. Kidney Int. 66:597–604. 2004. View Article : Google Scholar : PubMed/NCBI
25	Huang YJ, Mei YM, Wang YQ, et al: Transfection and expression of Smad7 inhibits transforming growth factor-β1 effects on renal tubular cells (Chinese). J Third Military Medical University. 25:1049–1052. 2003.