MicroRNA-129 Promotes Cardiomyogenesis in Bone Marrow Mesenchymal Stem Cells_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

LU Zhexin , LIAN Feng , XUE Song , ZHU Hongsheng .

Department of Cardiovascular Surgery，Renji Hospital，Medical School of ShanghaiJiaotong University，Shanghai 200147，P.R.China;

Corresponding author：

Keywords：

Bone marrow mesenchymal stem cells; 　Myocardial differentiation; 　MicroRNA-129; 　Glycogen synthase kinase-3β

DOI：

10.7507/1007-4848.20130176

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To explore the induction of cardiomyogenesis of microRNA-129 （mir-129） in rat bone marrowmesenchymal stem cells （BM-MSCs） and its mechanism. Methods BM-MSCs were isolated from Sprague-Dawley rats and cultured in vitro. Overexpression of mir-129 or both mir-129 and glycogen synthase kinase-3β （GSK-3β） in BM-MSCs was produced with a lentiviral vector system. All the BM-MSCs were divided into four groups: control group （MSCs），Lentiviral vectors＋MSCs group （Lv-MSCs），mir-129 transfection group （mir-129-MSCs），and mir-129＋GSK-3βdouble transfection group （mir-129＋GSK-3β-MSCs）. Five-Azacytidine （5-Aza）（10 μmol/L） was used to induce BM-MSCsdifferentiation into cardiomyocytes. On the 1st，5 th，10 th，15 th and 20 th day after induction，realtime-PCR was performedto detect mRNA levels of GATA-4，Nkx2.5 and MEF-2C. On the 10 th，15 th and 20 th day after induction，Western blottingwas performed to examine expression levels of cTnI，Desmin，GSK-3β，phosphorylated β-catenin and dephosphorylated β-catenin. Results Compared with the control group，at respective time points，mRNA levels of cardiomyogenic genes and expression levels of cardiomyocyte-related proteins of mir-129 transfection group were significantly elevated，theexpression level of GSK-3β was significantly decreased，and the ratio of dephosphorylated/phosphorylated β-catenin was significantly elevated. When both mir-129 and GSK-3β were overexpressed in BM-MSCs，mRNA levels of cardiomyogenicgenes and expression levels of cardiomyocyte-related proteins were significantly lower than those of mir-129 transfection group，and the ratio of dephosphorylated/phosphorylated β-catenin was significantly decreased. Conclusion Overexpression of mir-129 can promote cardiomyogenesis of rat BM-MSCs possibly via inhibiting GSK-3β production and thus decreasing the inhibition of phosphorylation of β-catenin which then enters the nucleus and activates downstream signaling pathways that regulate cardiomyogenic differentiation of BM-MSCs.

Citation： LU Zhexin,LIAN Feng,XUE Song,ZHU Hongsheng .. MicroRNA-129 Promotes Cardiomyogenesis in Bone Marrow Mesenchymal Stem Cells. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2013, 20(5): 570-576. doi: 10.7507/1007-4848.20130176 Copy

1.	Gnecchi M， Danieli P， Cervio E. Mesenchymal stem cell therapy for heart disease. Vascul Pharmacol， 2012， 57 （1）：48-55.
2.	Pillai RS. MicroRNA function：multiple mechanisms for a tiny RNA ? RNA， 2005， 11 （12）：1753-1761.
3.	Zamore PD， Haley B. Ribo-gnome：the big world of small RNAs. Science， 2005， 309 （5740）：1519-1524.
4.	Fu JD， Rushing SN， Lieu DK， et al. Distinct roles of microRNA-1 and -499 in ventricular specification and functional maturation ofhuman embryonic stem cell-derived cardiomyocytes. PLoS One， 2011， 6 （11）：e27417.
5.	Sun Q， Zhang Y， Yang G， et al. Transforming growth factor-beta-regulated miR-24 promotes skeletal muscle differentiation. Nucleic Acids Res， 2008， 36 （8）：2690-2699.
6.	Townley-Tilson WH， Callis TE， Wang D. MicroRNAs 1， 133， and 206：critical factors of skeletal and cardiac muscle development， function， and disease. Int J Biochem Cell Biol， 2010， 42 （8）：1252-1255.
7.	Zhang LL， Liu JJ， Liu F， et al. MiR-499 induces cardiac differentiation of rat mesenchymal stem cells through wnt/β-catenin signaling pathway. Biochem Biophys Res Commun， 2012， 420 （4）：875-881.
8.	Brunt KR， Zhang Y， Mihic A， et al. Role of WNT/β-Catenin signaling in rejuvenating myogenic. Am J Pathol， 2012， 181 （6）：2067-2078.
9.	Carthy JM， Garmaroudi FS， Luo Z， et al. Wnt3a induces myofibro-blast differentiation by upregulating TGF-β signaling through SMAD2in a β-catenin-dependent manner. PLoS One， 2011， 6 （5）：e19809.
10.	Blankesteijn WM， van de Schans VA， ter Horst P， et al. The Wnt/frizzled/GSK-3 beta pathway：a novel therapeutic target for cardiac hypertrophy. Trends Pharmacol Sci， 2008， 29 （4）：175-180.
11.	Liu Y， Huang T， Zhao X， et al. MicroRNAs modulate the Wnt signaling pathway through targeting its inhibitors. Biochem Biophys Res Commun， 2011， 408 （2）：259-264.
12.	Traverse JH， Henry TD， Moye L. Is the measurement of left ventr-icular ejection fraction the proper end point for cell therapy trials ? An analysis of the effect of bone marrow mononuclear stem cell administration on left ventricular ejection fraction after ST-segment elevation myocardial infarction when evaluated by cardiac magnetic resonance imaging. Am Heart J， 2011， 162 （4）：671-677.
13.	Britten MB， Abolmaali ND， Assmus B， et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction （TOPCARE-AMI）：mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation， 2003， 108 （18）：2212-2218.
14.	Rosca AM， Burlacu A. Effect of 5-azacytidine：evidence for alteration of the multipotent ability of mesenchymal stem cells. Stem Cells Dev， 2011， 20 （7）：1213-1221.
15.	Ramesh B， Bishi DK， Rallapalli S， et al. Ischemic cardiac tissue conditioned media induced differentiation of human mesenchymal stem cells into early stage cardiomyocytes. Cytotechnology， 2012， 64 （5）：563-575.
16.	朱洪生，连锋，朱伟，等. 骨髓间质干细胞体外转化为心肌细胞的实验研究. 上海第二医科大学学报， 2002， 22 （5）：391-395.
17.	Katoh M， Katoh M. WNT signaling pathway and stem cell signaling network. Clin Cancer Res， 2007， 13 （14）：4042-4045.
18.	路喆鑫，连锋. 分泌型卷曲相关蛋白2在心肌再生治疗中的作用. 中国胸心血管外科临床杂志， 2012， 19 （4）：423-426.
19.	Nelson WJ， Nusse R . Convergence of Wnt， beta-catenin， and cadherin pathways.Science， 2004， 303 （5663）：1483-1487.
20.	Zeng X， Tamai K， Doble B， et al. A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. Nature， 2005， 438 （7069）：873-877.
21.	Doble BW， Woodgett JR. GSK-3：tricks of the trade for a multi-taskingkinase. J Cell Sci， 2003， 116 （Pt 7）：1175-1186.
22.	Cho J， Rameshwar P， Sadoshima J. Distinct roles of glycogen synthasekinase （GSK）-3alpha and GSK-3beta in mediating cardiomyocyte differentiation in murine bone marrow-derived mesenchymal stem cells. J Biol Chem， 2009， 284 （52）：36647-36658.
23.	Kwon C， Cheng P， King IN， et al. Notch post-translationally regulates β-catenin protein in stem and progenitor cells. Nat Cell Biol， 2011，13 （10）：1244-1251.
24.	Jia J， Amanai K， Wang G， et al. Shaggy/GSK3 antagonizes Hedgehog signalling by regulating Cubitus interruptus. Nature， 2002， 416 （6880）：548-552.
25.	Hikasa H， Sokol SY. Phosphorylation of TCF proteins by homeodomain-interacting protein kinase 2. J Biol Chem， 2011， 286 （14）：12093-12100.

1. 　Gnecchi M， Danieli P， Cervio E. Mesenchymal stem cell therapy for heart disease. Vascul Pharmacol， 2012， 57 （1）：48-55.
2. 　Pillai RS. MicroRNA function：multiple mechanisms for a tiny RNA ? RNA， 2005， 11 （12）：1753-1761.
3. 　Zamore PD， Haley B. Ribo-gnome：the big world of small RNAs. Science， 2005， 309 （5740）：1519-1524.
4. 　Fu JD， Rushing SN， Lieu DK， et al. Distinct roles of microRNA-1 and -499 in ventricular specification and functional maturation ofhuman embryonic stem cell-derived cardiomyocytes. PLoS One， 2011， 6 （11）：e27417.
5. 　Sun Q， Zhang Y， Yang G， et al. Transforming growth factor-beta-regulated miR-24 promotes skeletal muscle differentiation. Nucleic Acids Res， 2008， 36 （8）：2690-2699.
6. 　Townley-Tilson WH， Callis TE， Wang D. MicroRNAs 1， 133， and 206：critical factors of skeletal and cardiac muscle development， function， and disease. Int J Biochem Cell Biol， 2010， 42 （8）：1252-1255.
7. 　Zhang LL， Liu JJ， Liu F， et al. MiR-499 induces cardiac differentiation of rat mesenchymal stem cells through wnt/β-catenin signaling pathway. Biochem Biophys Res Commun， 2012， 420 （4）：875-881.
8. 　Brunt KR， Zhang Y， Mihic A， et al. Role of WNT/β-Catenin signaling in rejuvenating myogenic. Am J Pathol， 2012， 181 （6）：2067-2078.
9. 　Carthy JM， Garmaroudi FS， Luo Z， et al. Wnt3a induces myofibro-blast differentiation by upregulating TGF-β signaling through SMAD2in a β-catenin-dependent manner. PLoS One， 2011， 6 （5）：e19809.
10. 　Blankesteijn WM， van de Schans VA， ter Horst P， et al. The Wnt/frizzled/GSK-3 beta pathway：a novel therapeutic target for cardiac hypertrophy. Trends Pharmacol Sci， 2008， 29 （4）：175-180.
11. 　Liu Y， Huang T， Zhao X， et al. MicroRNAs modulate the Wnt signaling pathway through targeting its inhibitors. Biochem Biophys Res Commun， 2011， 408 （2）：259-264.
12. 　Traverse JH， Henry TD， Moye L. Is the measurement of left ventr-icular ejection fraction the proper end point for cell therapy trials ? An analysis of the effect of bone marrow mononuclear stem cell administration on left ventricular ejection fraction after ST-segment elevation myocardial infarction when evaluated by cardiac magnetic resonance imaging. Am Heart J， 2011， 162 （4）：671-677.
13. 　Britten MB， Abolmaali ND， Assmus B， et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction （TOPCARE-AMI）：mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation， 2003， 108 （18）：2212-2218.
14. 　Rosca AM， Burlacu A. Effect of 5-azacytidine：evidence for alteration of the multipotent ability of mesenchymal stem cells. Stem Cells Dev， 2011， 20 （7）：1213-1221.
15. 　Ramesh B， Bishi DK， Rallapalli S， et al. Ischemic cardiac tissue conditioned media induced differentiation of human mesenchymal stem cells into early stage cardiomyocytes. Cytotechnology， 2012， 64 （5）：563-575.
16. 　朱洪生，连锋，朱伟，等. 骨髓间质干细胞体外转化为心肌细胞的实验研究. 上海第二医科大学学报， 2002， 22 （5）：391-395.
17. 　Katoh M， Katoh M. WNT signaling pathway and stem cell signaling network. Clin Cancer Res， 2007， 13 （14）：4042-4045.
18. 　路喆鑫，连锋. 分泌型卷曲相关蛋白2在心肌再生治疗中的作用. 中国胸心血管外科临床杂志， 2012， 19 （4）：423-426.
19. 　Nelson WJ， Nusse R . Convergence of Wnt， beta-catenin， and cadherin pathways.Science， 2004， 303 （5663）：1483-1487.
20. 　Zeng X， Tamai K， Doble B， et al. A dual-kinase mechanism for Wnt co-receptor phosphorylation and activation. Nature， 2005， 438 （7069）：873-877.
21. 　Doble BW， Woodgett JR. GSK-3：tricks of the trade for a multi-taskingkinase. J Cell Sci， 2003， 116 （Pt 7）：1175-1186.
22. 　Cho J， Rameshwar P， Sadoshima J. Distinct roles of glycogen synthasekinase （GSK）-3alpha and GSK-3beta in mediating cardiomyocyte differentiation in murine bone marrow-derived mesenchymal stem cells. J Biol Chem， 2009， 284 （52）：36647-36658.
23. 　Kwon C， Cheng P， King IN， et al. Notch post-translationally regulates β-catenin protein in stem and progenitor cells. Nat Cell Biol， 2011，13 （10）：1244-1251.
24. 　Jia J， Amanai K， Wang G， et al. Shaggy/GSK3 antagonizes Hedgehog signalling by regulating Cubitus interruptus. Nature， 2002， 416 （6880）：548-552.
25. 　Hikasa H， Sokol SY. Phosphorylation of TCF proteins by homeodomain-interacting protein kinase 2. J Biol Chem， 2011， 286 （14）：12093-12100.

Previous Article
Research Progression of Perineural Invasion of Tumor
Next Article
Research Progression of Zinc Deficiency after Gastric Bypass Surgery

Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

MicroRNA-129 Promotes Cardiomyogenesis in Bone Marrow Mesenchymal Stem Cells

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content