Mesenchymal Stem Cells Attenuates TGF-β1-Induced EMT by Increasing HGF Expression in HK-2 Cells
-
Junjun Wei
,
Li Tang
,
Liang-Liang Chen
,
Zhen-Hua Xie
,
Yu Ren
,
Hong-Gang Qi
,
Jiang-Yong Lou
,
Guo-Bin Weng
,
Shu-Wei ZHANG
Abstract
Background: Mesenchymal stem cells (MSCs) have recently shown promise for the treatment of various types of chronic kidney disease models. However, the mechanism of this effect is still not well understood. Our study is aimed to investigate the effect of MSCs on transforming growth factor beta 1 (TGF-β1)-induced epithelial mesenchymal transition (EMT) in renal tubular epithelial cells (HK-2 cells) and the underlying mechanism related to the reciprocal balance between hepatocyte growth factor (HGF) and TGF-β1.
Methods: Our study was performed at Ningbo University, Ningbo, Zhejiang, China between Mar 2017 and Jun 2018. HK-2 cells were initially treated with TGF-β1，then co-cultured with MSCs. The induced EMT was assessed by cellular morphology and the expressions of alpha-smooth muscle actin (α-SMA) and EMT-related proteins. MTS assay and flow cytometry were employed to detect the effect of TGF-β1 and MSCs on HK-2 cell proliferation and apoptosis. SiRNA against hepatocyte growth factor (siHGF) was transfected to decrease the expression of HGF to identify the role of HGF in MSCs inhibiting HK-2 cells EMT.
Results: Overexpressing TGF-β1 decreased HGF expression, induced EMT, suppressed proliferation and promoted apoptosis in HK-2 cells; but when co-cultured with MSCs all the outcomes were reversed. However, after treated with siHGF, all the benefits taken from MSCs vanished.
Conclusion: TGF-β1 was a motivating factor of kidney cell EMT and it suppressed the HGF expression. However, MSCs provided protection against EMT by increasing HGF level and decreasing TGF-β1 level. Our results also demonstrated HGF is one of the critical factor in MSCs anti- fibrosis.
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2. Uccelli A, Moretta L and Pistoia V (2008). Mesenchymal stem cells in health and disease. Nat Rev Immunol, 8(9): 726-36.
3. Asanuma H, Vanderbrink BA, Campbell MT, et al (2011). Arterially delivered mes-enchymal stem cells prevent obstruction-induced renal fibrosis. J Surg Res, 168(1): e51-9.
4. Caplan AI and Dennis JE (2006). Mesen-chymal stem cells as trophic mediators. J Cell Biochem, 98(5): 1076-84.
5. Jiang Z, Liu G, Meng F, et al (2017). Para-crine effects of mesenchymal stem cells on the activation of keratocytes. Br J Oph-thalmol, 101(11): 1583-90.
6. Gallo S, Sala V, Gatti S, et al (2015). Cellular and molecular mechanisms of HGF/Met in the cardiovascular system. Clin Sci (Lond), 129(12): 1173-93.
7. Hu ZP, Bao Y, Chen DN, et al (2013). Ef-fects of recombinant adenovirus hepato-cyte growth factor gene on myocardial remodeling in spontaneously hyperten-sive rats. J Cardiovasc Pharmacol Ther, 18(5): 476-80.
8. Wang Z, Fei S, Suo C, et al (2018). Antifi-brotic Effects of Hepatocyte Growth Factor on Endothelial-to-Mesenchymal Transition via Transforming Growth Factor-Beta1 (TGF-beta1)/Smad and Akt/mTOR/P70S6K Signaling Pathways. Ann Transplant, 23:1-10.
9. Border WA and Noble NA (1994). Trans-forming growth factor beta in tissue fi-brosis. N Engl J Med, 331(19): 1286-92.
10. Sakai K, Jawaid S, Sasaki T, et al (2014). Transforming growth factor-beta-independent role of connective tissue growth factor in the development of liver fibrosis. Am J Pathol, 184(10): 2611-7.
11. Guo L, Peng W, Tao J, et al (2016). Hydro-gen Sulfide Inhibits Transforming Growth Factor-beta1-Induced EMT via Wnt/Catenin Pathway. PLoS One, 11(1): e0147018.
12. Xu Y, Lou Z and Lee SH (2017). Arctigenin represses TGF-beta-induced epithelial mesenchymal transition in human lung cancer cells. Biochem Biophys Res Commun, 493(2): 934-39.
13. Shiota M, Zardan A, Takeuchi A, et al (2012). Clusterin mediates TGF-beta-induced epithelial-mesenchymal transition and metastasis via Twist1 in prostate can-cer cells. Cancer Res, 72(20): 5261-72.
14. Chen Y, Zhang K, Li Y, et al (2017). Estro-gen-related receptor alpha participates transforming growth factor-beta (TGF-beta) induced epithelial-mesenchymal transition of osteosarcoma cells. Cell Adh Migr, 11(4): 338-46.
15. Taiyab A, Korol A, Deschamps PA, et al (2016). beta-Catenin/CBP-Dependent Signaling Regulates TGF-beta-Induced Epithelial to Mesenchymal Transition of Lens Epithelial Cells. Invest Ophthalmol Vis Sci, 57(13): 5736-47.
16. Biname F, Lassus P and Hibner U (2008). Transforming growth factor beta con-trols the directional migration of hepato-cyte cohorts by modulating their adhe-sion to fibronectin. Mol Biol Cell, 19(3): 945-56.
17. Chattopadhyay N, J TFH, Godbole MM, et al (2004). Transforming growth factor beta receptor family ligands inhibit hepatocyte growth factor synthesis and secretion from astrocytoma cells. Brain Res Mol Brain Res, 121(1-2): 146-50.
18. Mungunsukh O, Day RM (2013). Trans-forming growth factor-beta1 selectively inhibits hepatocyte growth factor expres-sion via a micro-RNA-199-dependent posttranscriptional mechanism. Mol Biol Cell, 24(13): 2088-97.
19. Mizuno S, Matsumoto K, Kurosawa T, et al (2000). Reciprocal balance of hepatocyte growth factor and transforming growth factor-beta 1 in renal fibrosis in mice. Kidney Int, 57(3): 937-48.
20. Sutton MT, Fletcher D, Episalla N, et al (2017). Mesenchymal Stem Cell Soluble Mediators and Cystic Fibrosis. J Stem Cell Res Ther, 7(9):400.
21. Cheng GS, Zhang YS, Zhang TT, et al (2017). Bone marrow-derived mesen-chymal stem cells modified with IGFBP-3 inhibit the proliferation of pulmonary artery smooth muscle cells. Int J Mol Med, 39(1): 223-30.
22. Wang X, Ren Y, Zhuang H, et al (2015). Decrease of phosphorylated proto-oncogene CREB at Ser 133 site inhibits growth and metastatic activity of renal cell cancer. Expert Opin Ther Targets, 19(7): 985-95.
23. Liu T, Peng YF, Jia C, et al (2015). Effect of HGF on the apoptosis of rat corpus cav-ernosum smooth muscle cells induced by TGFbeta1. Andrologia, 47(9): 1020-7.
24. Guo HL, Liao XH, Liu Q, et al (2016). An-giotensin II Type 2 Receptor Decreases Transforming Growth Factor-beta Type II Receptor Expression and Function in Human Renal Proximal Tubule Cells. PLoS One, 11(2): e0148696.
25. Aghajani Nargesi A, Lerman LO and Eirin A (2017). Mesenchymal stem cell-derived extracellular vesicles for kidney repair: current status and looming challenges. Stem Cell Res Ther, 8(1): 273.
26. Fleig SV and Humphreys BD (2014). Ra-tionale of mesenchymal stem cell therapy in kidney injury. Nephron Clin Pract, 127(1-4): 75-80.
27. Serini G, Bochaton-Piallat ML, Ropraz P, et al (1998). The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-beta1. J Cell Biol, 142(3): 873-81.
28. Wang Z, Han Z, Tao J, et al (2017). Role of endothelial-to-mesenchymal transition induced by TGF-beta1 in transplant kid-ney interstitial fibrosis. J Cell Mol Med, 21(10): 2359-69.
29. Liu J, Wang F, Xie M, et al (2018). Response to inhibition of TGF-beta1 might be a novel therapeutic target in the treatment of cardiac fibrosis. Int J Cardiol, 256:20.
30. Tian XR, Tian XL, Wang HF, et al (2016). [Regulation effect of beta-catenin pathway on TGF-beta1 induced pulmonary pro-fibrosis]. Zhonghua Yi Xue Za Zhi, 96(24): 1929-33.
31. Kim KH, Kim HC, Hwang MY, et al (2006). The antifibrotic effect of TGF-beta1 siRNAs in murine model of liver cirrho-sis. Biochem Biophys Res Commun, 343(4): 1072-8.
32. Zhou B, Wang Y, Zhang C, et al (2018). Ri-bemansides A and B, TRPC6 Inhibitors from Ribes manshuricum That Suppress TGF-beta1-Induced Fibrogenesis in HK-2 Cells. J Nat Prod, 81(4): 913-17.
33. Jang YO, Cho MY, Yun CO, et al (2016). Effect of Function-Enhanced Mesen-chymal Stem Cells Infected With Decorin-Expressing Adenovirus on He-patic Fibrosis. Stem Cells Transl Med, 5(9): 1247-56.
34. Kennelly H, Mahon BP and English K (2016). Human mesenchymal stromal cells exert HGF dependent cytoprotective effects in a human relevant pre-clinical model of COPD. Sci Rep, 6:38207.
35. Massague J (2012). TGFbeta signalling in context. Nat Rev Mol Cell Biol, 13(10): 616-30.
36. Grande JP, Warner GM, Walker HJ, et al (2002). TGF-beta1 is an autocrine media-tor of renal tubular epithelial cell growth and collagen IV production. Exp Biol Med (Maywood), 227(3): 171-81.
37. Roushandeh AM, Bahadori M and Roudke-nar MH (2017). Mesenchymal Stem Cell-based Therapy as a New Horizon for Kidney Injuries. Arch Med Res, 48(2): 133-46.
38. Eliopoulos N, Zhao J, Bouchentouf M, et al (2010). Human marrow-derived mesen-chymal stromal cells decrease cisplatin renotoxicity in vitro and in vivo and en-hance survival of mice post-intraperitoneal injection. Am J Physiol Renal Physiol, 299(6): F1288-98.
39. Nakamura T, Sakai K, Nakamura T, et al (2011). Hepatocyte growth factor twenty years on: Much more than a growth fac-tor. J Gastroenterol Hepatol, 26 Suppl 1:188-202.
40. Wang H, Zheng R, Chen Q, et al (2017). Mesenchymal stem cells microvesicles stabilize endothelial barrier function partly mediated by hepatocyte growth factor (HGF). Stem Cell Res Ther, 8(1): 211.
41. Li W, Jiang H and Feng JM (2012). Isogenic mesenchymal stem cells transplantation improves a rat model of chronic aris-tolochic acid nephropathy via upregula-tion of hepatic growth factor and down-regulation of transforming growth factor beta1. Mol Cell Biochem, 368(1-2): 137-45.
42. Lu F, Zhao X, Wu J, et al (2013). MSCs transfected with hepatocyte growth factor or vascular endothelial growth factor im-prove cardiac function in the infarcted porcine heart by increasing angiogenesis and reducing fibrosis. Int J Cardiol, 167(6): 2524-32.
43. Lee EJ, Hwang I, Lee JY, et al (2018). Hepatocyte Growth Factor Improves the Therapeutic Efficacy of Human Bone Marrow Mesenchymal Stem Cells via RAD51. Mol Ther, 26(3): 845-59.
44. Halabian R, Roudkenar MH, Jahanian-Najafabadi A, et al (2015). Co-culture of bone marrow-derived mesenchymal stem cells overexpressing lipocalin 2 with HK-2 and HEK293 cells protects the kidney cells against cisplatin-induced injury. Cell Biol Int, 39(2): 152-63.
2. Uccelli A, Moretta L and Pistoia V (2008). Mesenchymal stem cells in health and disease. Nat Rev Immunol, 8(9): 726-36.
3. Asanuma H, Vanderbrink BA, Campbell MT, et al (2011). Arterially delivered mes-enchymal stem cells prevent obstruction-induced renal fibrosis. J Surg Res, 168(1): e51-9.
4. Caplan AI and Dennis JE (2006). Mesen-chymal stem cells as trophic mediators. J Cell Biochem, 98(5): 1076-84.
5. Jiang Z, Liu G, Meng F, et al (2017). Para-crine effects of mesenchymal stem cells on the activation of keratocytes. Br J Oph-thalmol, 101(11): 1583-90.
6. Gallo S, Sala V, Gatti S, et al (2015). Cellular and molecular mechanisms of HGF/Met in the cardiovascular system. Clin Sci (Lond), 129(12): 1173-93.
7. Hu ZP, Bao Y, Chen DN, et al (2013). Ef-fects of recombinant adenovirus hepato-cyte growth factor gene on myocardial remodeling in spontaneously hyperten-sive rats. J Cardiovasc Pharmacol Ther, 18(5): 476-80.
8. Wang Z, Fei S, Suo C, et al (2018). Antifi-brotic Effects of Hepatocyte Growth Factor on Endothelial-to-Mesenchymal Transition via Transforming Growth Factor-Beta1 (TGF-beta1)/Smad and Akt/mTOR/P70S6K Signaling Pathways. Ann Transplant, 23:1-10.
9. Border WA and Noble NA (1994). Trans-forming growth factor beta in tissue fi-brosis. N Engl J Med, 331(19): 1286-92.
10. Sakai K, Jawaid S, Sasaki T, et al (2014). Transforming growth factor-beta-independent role of connective tissue growth factor in the development of liver fibrosis. Am J Pathol, 184(10): 2611-7.
11. Guo L, Peng W, Tao J, et al (2016). Hydro-gen Sulfide Inhibits Transforming Growth Factor-beta1-Induced EMT via Wnt/Catenin Pathway. PLoS One, 11(1): e0147018.
12. Xu Y, Lou Z and Lee SH (2017). Arctigenin represses TGF-beta-induced epithelial mesenchymal transition in human lung cancer cells. Biochem Biophys Res Commun, 493(2): 934-39.
13. Shiota M, Zardan A, Takeuchi A, et al (2012). Clusterin mediates TGF-beta-induced epithelial-mesenchymal transition and metastasis via Twist1 in prostate can-cer cells. Cancer Res, 72(20): 5261-72.
14. Chen Y, Zhang K, Li Y, et al (2017). Estro-gen-related receptor alpha participates transforming growth factor-beta (TGF-beta) induced epithelial-mesenchymal transition of osteosarcoma cells. Cell Adh Migr, 11(4): 338-46.
15. Taiyab A, Korol A, Deschamps PA, et al (2016). beta-Catenin/CBP-Dependent Signaling Regulates TGF-beta-Induced Epithelial to Mesenchymal Transition of Lens Epithelial Cells. Invest Ophthalmol Vis Sci, 57(13): 5736-47.
16. Biname F, Lassus P and Hibner U (2008). Transforming growth factor beta con-trols the directional migration of hepato-cyte cohorts by modulating their adhe-sion to fibronectin. Mol Biol Cell, 19(3): 945-56.
17. Chattopadhyay N, J TFH, Godbole MM, et al (2004). Transforming growth factor beta receptor family ligands inhibit hepatocyte growth factor synthesis and secretion from astrocytoma cells. Brain Res Mol Brain Res, 121(1-2): 146-50.
18. Mungunsukh O, Day RM (2013). Trans-forming growth factor-beta1 selectively inhibits hepatocyte growth factor expres-sion via a micro-RNA-199-dependent posttranscriptional mechanism. Mol Biol Cell, 24(13): 2088-97.
19. Mizuno S, Matsumoto K, Kurosawa T, et al (2000). Reciprocal balance of hepatocyte growth factor and transforming growth factor-beta 1 in renal fibrosis in mice. Kidney Int, 57(3): 937-48.
20. Sutton MT, Fletcher D, Episalla N, et al (2017). Mesenchymal Stem Cell Soluble Mediators and Cystic Fibrosis. J Stem Cell Res Ther, 7(9):400.
21. Cheng GS, Zhang YS, Zhang TT, et al (2017). Bone marrow-derived mesen-chymal stem cells modified with IGFBP-3 inhibit the proliferation of pulmonary artery smooth muscle cells. Int J Mol Med, 39(1): 223-30.
22. Wang X, Ren Y, Zhuang H, et al (2015). Decrease of phosphorylated proto-oncogene CREB at Ser 133 site inhibits growth and metastatic activity of renal cell cancer. Expert Opin Ther Targets, 19(7): 985-95.
23. Liu T, Peng YF, Jia C, et al (2015). Effect of HGF on the apoptosis of rat corpus cav-ernosum smooth muscle cells induced by TGFbeta1. Andrologia, 47(9): 1020-7.
24. Guo HL, Liao XH, Liu Q, et al (2016). An-giotensin II Type 2 Receptor Decreases Transforming Growth Factor-beta Type II Receptor Expression and Function in Human Renal Proximal Tubule Cells. PLoS One, 11(2): e0148696.
25. Aghajani Nargesi A, Lerman LO and Eirin A (2017). Mesenchymal stem cell-derived extracellular vesicles for kidney repair: current status and looming challenges. Stem Cell Res Ther, 8(1): 273.
26. Fleig SV and Humphreys BD (2014). Ra-tionale of mesenchymal stem cell therapy in kidney injury. Nephron Clin Pract, 127(1-4): 75-80.
27. Serini G, Bochaton-Piallat ML, Ropraz P, et al (1998). The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-beta1. J Cell Biol, 142(3): 873-81.
28. Wang Z, Han Z, Tao J, et al (2017). Role of endothelial-to-mesenchymal transition induced by TGF-beta1 in transplant kid-ney interstitial fibrosis. J Cell Mol Med, 21(10): 2359-69.
29. Liu J, Wang F, Xie M, et al (2018). Response to inhibition of TGF-beta1 might be a novel therapeutic target in the treatment of cardiac fibrosis. Int J Cardiol, 256:20.
30. Tian XR, Tian XL, Wang HF, et al (2016). [Regulation effect of beta-catenin pathway on TGF-beta1 induced pulmonary pro-fibrosis]. Zhonghua Yi Xue Za Zhi, 96(24): 1929-33.
31. Kim KH, Kim HC, Hwang MY, et al (2006). The antifibrotic effect of TGF-beta1 siRNAs in murine model of liver cirrho-sis. Biochem Biophys Res Commun, 343(4): 1072-8.
32. Zhou B, Wang Y, Zhang C, et al (2018). Ri-bemansides A and B, TRPC6 Inhibitors from Ribes manshuricum That Suppress TGF-beta1-Induced Fibrogenesis in HK-2 Cells. J Nat Prod, 81(4): 913-17.
33. Jang YO, Cho MY, Yun CO, et al (2016). Effect of Function-Enhanced Mesen-chymal Stem Cells Infected With Decorin-Expressing Adenovirus on He-patic Fibrosis. Stem Cells Transl Med, 5(9): 1247-56.
34. Kennelly H, Mahon BP and English K (2016). Human mesenchymal stromal cells exert HGF dependent cytoprotective effects in a human relevant pre-clinical model of COPD. Sci Rep, 6:38207.
35. Massague J (2012). TGFbeta signalling in context. Nat Rev Mol Cell Biol, 13(10): 616-30.
36. Grande JP, Warner GM, Walker HJ, et al (2002). TGF-beta1 is an autocrine media-tor of renal tubular epithelial cell growth and collagen IV production. Exp Biol Med (Maywood), 227(3): 171-81.
37. Roushandeh AM, Bahadori M and Roudke-nar MH (2017). Mesenchymal Stem Cell-based Therapy as a New Horizon for Kidney Injuries. Arch Med Res, 48(2): 133-46.
38. Eliopoulos N, Zhao J, Bouchentouf M, et al (2010). Human marrow-derived mesen-chymal stromal cells decrease cisplatin renotoxicity in vitro and in vivo and en-hance survival of mice post-intraperitoneal injection. Am J Physiol Renal Physiol, 299(6): F1288-98.
39. Nakamura T, Sakai K, Nakamura T, et al (2011). Hepatocyte growth factor twenty years on: Much more than a growth fac-tor. J Gastroenterol Hepatol, 26 Suppl 1:188-202.
40. Wang H, Zheng R, Chen Q, et al (2017). Mesenchymal stem cells microvesicles stabilize endothelial barrier function partly mediated by hepatocyte growth factor (HGF). Stem Cell Res Ther, 8(1): 211.
41. Li W, Jiang H and Feng JM (2012). Isogenic mesenchymal stem cells transplantation improves a rat model of chronic aris-tolochic acid nephropathy via upregula-tion of hepatic growth factor and down-regulation of transforming growth factor beta1. Mol Cell Biochem, 368(1-2): 137-45.
42. Lu F, Zhao X, Wu J, et al (2013). MSCs transfected with hepatocyte growth factor or vascular endothelial growth factor im-prove cardiac function in the infarcted porcine heart by increasing angiogenesis and reducing fibrosis. Int J Cardiol, 167(6): 2524-32.
43. Lee EJ, Hwang I, Lee JY, et al (2018). Hepatocyte Growth Factor Improves the Therapeutic Efficacy of Human Bone Marrow Mesenchymal Stem Cells via RAD51. Mol Ther, 26(3): 845-59.
44. Halabian R, Roudkenar MH, Jahanian-Najafabadi A, et al (2015). Co-culture of bone marrow-derived mesenchymal stem cells overexpressing lipocalin 2 with HK-2 and HEK293 cells protects the kidney cells against cisplatin-induced injury. Cell Biol Int, 39(2): 152-63.
| Files | ||
| Issue | Vol 50 No 5 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v50i5.6108 | |
| Keywords | ||
| Mesenchymal stem cells Transforming growth factor beta 1 (TGF-β1) Epithelial mesenchymal transition (EMT) Hepatocyte growth factor (HGF) Apoptosis | ||
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How to Cite
1.
Wei J, Tang L, Chen L-L, Xie Z-H, Ren Y, Qi H-G, Lou J-Y, Weng G-B, ZHANG S-W. Mesenchymal Stem Cells Attenuates TGF-β1-Induced EMT by Increasing HGF Expression in HK-2 Cells. Iran J Public Health. 2021;50(5):908-918.
