Effect of up-regulation of miRNA-21 in vitro on bone marrow mesenchymal stem cells_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

YANG Wengang ,  XUE Song , WEN Dezhong , ZHENG Hui , SHAN Jianggui

Department of Cardiovascular Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, P.R.China;

Corresponding author：

XUE Song, Email: xuesong64@163.com

Keywords：

Bone marrow mesenchymal stem cells; miRNA-21; proliferation; apoptosis; differentiation

DOI：

10.7507/1007-4848.201909021

Video：

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Objective To explore the effect of expression of miRNA-21 on bone marrow mesenchymal stem cells (BMSCs).Methods In this study, flow cytometry was used to identify the surface-associated antigens of BMSCs. The 10 μmol/L 5-azacytidine was used to induce BMSCs to differentiate to cardiomyocyte-like cells. Immunofluorescence was used to detect the expression of troponin I (cTnI). The samples were assigned to 3 groups: a blank group, a miRNA-21 mimic group, and a negative control (NC) group. The proliferation of BMSCs was detected by methyl thiazolylte-trazolium (MTT), the apoptosis of BMSCs was analyzed by flow cytometry. Western-blotting was used to identify the expression of cTnI and myod in the BMSCs.Results The proliferation of BMSCs was increased, because of the over expression of miRNA-21. But the apoptotic rate of the BMSCs was slower in the miRNA-21 group, on account of the expression of miRNA-21 was higher than that in the NC group and the CK group. The expression of cTnI in the miRNA-21 group was higher than that in the NC group or the CK group.Conclusion The results suggest that the up-regulation of miRNA-21 enhances proliferation of BMSCs, reduces the apoptosis of BMSCs. miRNA-21 promotes the differentiation of BMSCs, which may pave the way for the treatment directed toward restoring miRNA-21 function for myocardial ischemia.

Citation： YANG Wengang, XUE Song, WEN Dezhong, ZHENG Hui, SHAN Jianggui. Effect of up-regulation of miRNA-21 in vitro on bone marrow mesenchymal stem cells. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2020, 27(8): 940-947. doi: 10.7507/1007-4848.201909021 Copy

1.	Beyer Nardi N, da Silva Meirelles L. Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handb Exp Pharmacol, 2006, (174): 249-282.
2.	Brighton CT, Hunt RM. Early histological and ultrastructural changes in medullary fracture callus. J Bone Joint Surg Am, 1991, 73: 832-847.
3.	Minguell JJ, Erices A, Conget P. Mesenchymal stem cells. Exp Biol Med (Maywood), 2001, 226(6): 507-520.
4.	Sensebé L, Krampera M, Schrezenmeier H, et al. Mesenchymal stem cells for clinical application. Vox Sang, 2010, 98(2): 93-107.
5.	Wang W, Jiang Q, Zhang H, et al. Intravenous administration of bone marrow mesenchymal stromal cells is safe for the lung in a chronic myocardial infarction model. Regen Med, 2011, 6(2): 179-190.
6.	Miyahara Y, Nagaya N, Kataoka M, et al. Monolayered mesen-chymal stem cells repair scarred myocardium after myocardial infarction. Nat Med, 2006, 12: 459-465.
7.	Nagaya N, Kangawa K, Itoh T, et al. Transplantation of mesen-chymal stem cells improves cardiac function in a rat model of dilated cardiomyopathy. Circulation, 2005, 112: 1128-1135.
8.	Eguchi T, Watanabe K, Hara ES, et al. OstemiR: a novel panel of microRNA biomarkers in osteoblastic and osteocytic differentiation from mesencymal stem cells. PLoS One, 2013, 8(3): e58796.
9.	Fakhry M, Hamade E, Badran B, et al. Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts. World J Stem Cells, 2013, 5: 136-148.
10.	Lakshmipathy U, Hart RP. Concise review: microRNA expression in multipotent mesenchymal stromal cells. Stem Cells, 2008, 26: 356-363.
11.	Sekar D, Hairul Islam VI, Thirugnanasambantham K, et al. Relevance of miR-21 in HIV and non-HIV-related lymphomas. Tumour Biol, 2014, 35(9): 8387-8393.
12.	Mathieu J, Ruohola-Baker H. Regulation of stem cell populations by microRNAs. Adv Exp Med Biol, 2013, 786: 329-351.
13.	Zou DD, Tang XB. Epithelial-mesenchymal transition and cancer stem cells. Zhonghua Bing Li Xue Za Zhi, 2013, 42: 62-65.
14.	Zhang R, Wang D, Xia Z, et al. The role of microRNAs in adipocyte differentiation. Front Med, 2013, 7: 223-230.
15.	Ceppi P, Peter ME. MicroRNAs regulate both epithelial-to-mesenchymal transition and cancer stem cells. Oncogene, 2014, 33(3): 269-278.
16.	Kang HY. MicroRNA-21 regulates stemness in cancer cells. Stem Cell Res Ther, 2013, 4(5): 110.
17.	Bakhshandeh B, Hafizi M, Ghaemi N, et al. Down-regulation of miRNA-221 triggers osteogenic differentiation in human stem cells. Biotechnol Lett, 2012, 34(8): 1579-1587.
18.	Trohatou O, Zagoura D, Bitsika V, et al. Sox2 suppression by miR-21 governs human mesenchymal stem cell properties. Stem Cells Transl Med, 2014, 3(1): 54-68.
19.	Mei Y, Bian C, Li J, et al. miR-21 modulates the ERK-MAPK signaling pathway by regulating SPRY2 expression during human mesenchymal stem cell differentiation. J Cell Biochem, 2013, 114(6): 1374-1384.
20.	Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defning multipotent mesenchymal stromal cells. The International Society for Cellular Therapy Position statement. Cytotherapy, 2006, 8: 315-317.
21.	Friedenstein AJ, Petrakova KV, Kurolesova AI, et al. Heterotopic transplants of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation, 1968, 6: 230-247.
22.	Dennis JE, Charbord P. Origin and differentiation of human and murine stroma. Stem Cells, 2002, 20(3): 205-214.
23.	Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood, 2001, 98(8): 2396-2402.
24.	Martin DR, Cox NR, Hathcock TL, et al. Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol, 2002, 30(8): 879-886.
25.	Wakitani S, Saito T and Caplan AI. Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve, 1995, 18: 1417-1426.
26.	Xu W, Zhang X, Qian H, et al. Mesenchymal stem cells from adult human bone marrow differentiate into a cardiomyocyte phenotype in vitro. Exp Biol Med (Maywood), 2004, 229: 623-631.
27.	Makino S, Fukuda K, Miyoshi S, et al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest, 1999, 103(5): 697-705.
28.	Toma C, Pittenger MF, Cahill KS, et al. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation, 2002, 105(1): 93-98.
29.	Yoon J, Min BG, Kim YH, et al. Differentiation, engraftment and functional effects of pre-treated mesenchymal stem cells in a rat myocardial infarct model. Acta Cardiol, 2005, 60(3): 277-284.
30.	Xie XJ, Wang JA, Cao J, et al. Differentiation of bone marrow mesenchymal stem cells induced by myocardial medium under hypoxic conditions. Acta Pharmacol Sin, 2006, 27: 1153-1158.
31.	Forte G, Minieri M, Cossa P, et al. Hepatocyte growth factor effects on mesenchymal stem cells: proliferation, migration, and differentiation. Stem Cells, 2006, 24(1): 23-33.
32.	Yang N, Wang G, Hu C, et al. Tumor necrosis factor α suppresses the mesenchymal stem cell osteogenesis promoter miR-21 in estrogen deficiency-induced osteoporosis. J Bone Miner Res, 2013, 28(3): 559-573.
33.	Zhang Y, Jia J, Yang S, et al. MicroRNA-21 controls the development of osteoarthritis by targeting GDF-5 in chondrocytes. Exp Mol Med, 2014, 46: e79.
34.	Shin KK, Lee AL, Kim JY, et al. miR-21 modulates tumor outgrowth induced by human adipose tissue-derived mesenchymal stem cells in vivo. Biochem Biophys Res Commun, 2012, 422(4): 633-638.
35.	Sekar D, Saravanan S, Karikalan K, et al. Role of microRNA 21 in mesenchymal stem cell (MSC) differentiation: a powerful biomarker in MSCs derived cells. Curr Pharm Biotechnol, 2015, 16(1): 43-48.

1. Beyer Nardi N, da Silva Meirelles L. Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handb Exp Pharmacol, 2006, (174): 249-282.
2. Brighton CT, Hunt RM. Early histological and ultrastructural changes in medullary fracture callus. J Bone Joint Surg Am, 1991, 73: 832-847.
3. Minguell JJ, Erices A, Conget P. Mesenchymal stem cells. Exp Biol Med (Maywood), 2001, 226(6): 507-520.
4. Sensebé L, Krampera M, Schrezenmeier H, et al. Mesenchymal stem cells for clinical application. Vox Sang, 2010, 98(2): 93-107.
5. Wang W, Jiang Q, Zhang H, et al. Intravenous administration of bone marrow mesenchymal stromal cells is safe for the lung in a chronic myocardial infarction model. Regen Med, 2011, 6(2): 179-190.
6. Miyahara Y, Nagaya N, Kataoka M, et al. Monolayered mesen-chymal stem cells repair scarred myocardium after myocardial infarction. Nat Med, 2006, 12: 459-465.
7. Nagaya N, Kangawa K, Itoh T, et al. Transplantation of mesen-chymal stem cells improves cardiac function in a rat model of dilated cardiomyopathy. Circulation, 2005, 112: 1128-1135.
8. Eguchi T, Watanabe K, Hara ES, et al. OstemiR: a novel panel of microRNA biomarkers in osteoblastic and osteocytic differentiation from mesencymal stem cells. PLoS One, 2013, 8(3): e58796.
9. Fakhry M, Hamade E, Badran B, et al. Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts. World J Stem Cells, 2013, 5: 136-148.
10. Lakshmipathy U, Hart RP. Concise review: microRNA expression in multipotent mesenchymal stromal cells. Stem Cells, 2008, 26: 356-363.
11. Sekar D, Hairul Islam VI, Thirugnanasambantham K, et al. Relevance of miR-21 in HIV and non-HIV-related lymphomas. Tumour Biol, 2014, 35(9): 8387-8393.
12. Mathieu J, Ruohola-Baker H. Regulation of stem cell populations by microRNAs. Adv Exp Med Biol, 2013, 786: 329-351.
13. Zou DD, Tang XB. Epithelial-mesenchymal transition and cancer stem cells. Zhonghua Bing Li Xue Za Zhi, 2013, 42: 62-65.
14. Zhang R, Wang D, Xia Z, et al. The role of microRNAs in adipocyte differentiation. Front Med, 2013, 7: 223-230.
15. Ceppi P, Peter ME. MicroRNAs regulate both epithelial-to-mesenchymal transition and cancer stem cells. Oncogene, 2014, 33(3): 269-278.
16. Kang HY. MicroRNA-21 regulates stemness in cancer cells. Stem Cell Res Ther, 2013, 4(5): 110.
17. Bakhshandeh B, Hafizi M, Ghaemi N, et al. Down-regulation of miRNA-221 triggers osteogenic differentiation in human stem cells. Biotechnol Lett, 2012, 34(8): 1579-1587.
18. Trohatou O, Zagoura D, Bitsika V, et al. Sox2 suppression by miR-21 governs human mesenchymal stem cell properties. Stem Cells Transl Med, 2014, 3(1): 54-68.
19. Mei Y, Bian C, Li J, et al. miR-21 modulates the ERK-MAPK signaling pathway by regulating SPRY2 expression during human mesenchymal stem cell differentiation. J Cell Biochem, 2013, 114(6): 1374-1384.
20. Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defning multipotent mesenchymal stromal cells. The International Society for Cellular Therapy Position statement. Cytotherapy, 2006, 8: 315-317.
21. Friedenstein AJ, Petrakova KV, Kurolesova AI, et al. Heterotopic transplants of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation, 1968, 6: 230-247.
22. Dennis JE, Charbord P. Origin and differentiation of human and murine stroma. Stem Cells, 2002, 20(3): 205-214.
23. Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood, 2001, 98(8): 2396-2402.
24. Martin DR, Cox NR, Hathcock TL, et al. Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol, 2002, 30(8): 879-886.
25. Wakitani S, Saito T and Caplan AI. Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve, 1995, 18: 1417-1426.
26. Xu W, Zhang X, Qian H, et al. Mesenchymal stem cells from adult human bone marrow differentiate into a cardiomyocyte phenotype in vitro. Exp Biol Med (Maywood), 2004, 229: 623-631.
27. Makino S, Fukuda K, Miyoshi S, et al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest, 1999, 103(5): 697-705.
28. Toma C, Pittenger MF, Cahill KS, et al. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation, 2002, 105(1): 93-98.
29. Yoon J, Min BG, Kim YH, et al. Differentiation, engraftment and functional effects of pre-treated mesenchymal stem cells in a rat myocardial infarct model. Acta Cardiol, 2005, 60(3): 277-284.
30. Xie XJ, Wang JA, Cao J, et al. Differentiation of bone marrow mesenchymal stem cells induced by myocardial medium under hypoxic conditions. Acta Pharmacol Sin, 2006, 27: 1153-1158.
31. Forte G, Minieri M, Cossa P, et al. Hepatocyte growth factor effects on mesenchymal stem cells: proliferation, migration, and differentiation. Stem Cells, 2006, 24(1): 23-33.
32. Yang N, Wang G, Hu C, et al. Tumor necrosis factor α suppresses the mesenchymal stem cell osteogenesis promoter miR-21 in estrogen deficiency-induced osteoporosis. J Bone Miner Res, 2013, 28(3): 559-573.
33. Zhang Y, Jia J, Yang S, et al. MicroRNA-21 controls the development of osteoarthritis by targeting GDF-5 in chondrocytes. Exp Mol Med, 2014, 46: e79.
34. Shin KK, Lee AL, Kim JY, et al. miR-21 modulates tumor outgrowth induced by human adipose tissue-derived mesenchymal stem cells in vivo. Biochem Biophys Res Commun, 2012, 422(4): 633-638.
35. Sekar D, Saravanan S, Karikalan K, et al. Role of microRNA 21 in mesenchymal stem cell (MSC) differentiation: a powerful biomarker in MSCs derived cells. Curr Pharm Biotechnol, 2015, 16(1): 43-48.

Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Effect of up-regulation of miRNA-21 in vitro on bone marrow mesenchymal stem cells

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