Biotechnology Letters

, Volume 38, Issue 4, pp 579–587 | Cite as

Restoration of miR-98 relieves the inhibitory effect of nicotine on the differentiation of P19 cells into cardiomyocytes

Original Research Paper
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Abstract

Objective

To study whether miR-98 participates in the effects of nicotine on myocardial differentiation.

Results

By western blot, MTT and flow cytometry assays, we found that nicotine suppresses P19 cell differentiation into cardiomyocytes and apoptosis, and promotes proliferation, while restoration of miR-98 relieves the inhibitory effect of nicotine on the P19 cell differentiation. By target prediction analysis and luciferase reporter assay, we observed that miR-98 inhibits the protein expression of Wnt1 by directly acting on the 3′-UTR of Wnt1 mRNA. We assumed that the effect of miR-98 on Wnt1 might alter the activity of the Wnt1/β-catenin signaling pathway and be associated with myocardial differentiation. In summary, nicotine restrains differentiation of P19 cells into cardiomyocytes and decreases the level of miR-98.

Conclusions

Restoration of miR-98 relieves the inhibitory effect of nicotine on differentiation of P19 cells via targeting the 3′-UTR of Wnt1, which offers novel insights into our understanding of underlying molecular mechanisms of congenital heart defects.

Keywords

Cardiomyocytes Congenital heart defect miR-98 Myocardial differentiation Nicotine P19 cells Wnt1 

Notes

Compliance with ethical standards

Conflict of interest

All authors declare that there is no conflict of interest.

References

  1. Cohen ED, Tian Y, Morrisey EE (2008) Wnt signaling: an essential regulator of cardiovascular differentiation, morphogenesis and progenitor self-renewal. Development 135:789–798CrossRefPubMedGoogle Scholar
  2. Grooms K, Hart M, Kang B, Wongtrakool C (2013) Mir-98 reduces nicotine-induced nerve growth factor (ngf) expression in murine lung fibroblasts. Am J Respir Crit Care Med 187:A3430Google Scholar
  3. Hu DL, Liu YQ, Chen FK, Sheng YH, Yang R et al (2011) Differential expression of microRNAs in cardiac myocytes compared to undifferentiated P19 cells. Int J Mol Med 28:59–64PubMedGoogle Scholar
  4. Kaczocha M, Glaser ST, Deutsch DG (2009) Identification of intracellular carriers for the endocannabinoid anandamide. Proc Natl Acad Sci USA 106:6375–6380CrossRefPubMedPubMedCentralGoogle Scholar
  5. Lévy M, Maurey C, Celermajer DS, Vouhé PR, Danel C, Bonnet D, Israël-Biet D (2007) Impaired apoptosis of pulmonary endothelial cells is associated with intimal proliferation and irreversibility of pulmonary hypertension in congenital heart disease. J Am Coll Cardiol 49:803–810CrossRefPubMedGoogle Scholar
  6. Li Q, Gregory RI (2008) MicroRNA regulation of stem cell fate. Cell Stem Cell 2:195–196CrossRefPubMedGoogle Scholar
  7. Ma K, Zhao Q, Chen W, Zhang H, Li S, Pan X, Chen Q (2015) Human lung microRNA profiling in pulmonary arterial hypertension secondary to congenital heart defect. Pediatr PulmonolGoogle Scholar
  8. Marvin MJ, Di Rocco G, Gardiner A, Bush SM, Lassar AB (2001) Inhibition of Wnt activity induces heart formation from posterior mesoderm. Genes Dev 15:316–327CrossRefPubMedPubMedCentralGoogle Scholar
  9. Miyazono K, Kamiya Y, Morikawa M (2010) Bone morphogenetic protein receptors and signal transduction. J Biochem 147:35–51CrossRefPubMedGoogle Scholar
  10. Qin DN, Qian L, Hu DL, Yu ZB, Han SP, Zhu C et al (2013) Effects of miR-19b overexpression on proliferation, differentiation, apoptosis and Wnt/β-catenin signaling pathway in P19 cell model of cardiac differentiation in vitro. Cell Biochem Biophys 66:709–722CrossRefPubMedGoogle Scholar
  11. Riehle-Colarusso T, Autry A, Razzaghi H, Boyle CA, Mahle WT, Braun KVN, Correa A (2015) Congenital heart defects and receipt of special education services. Pediatrics 136:496–504CrossRefPubMedPubMedCentralGoogle Scholar
  12. Shi M, Wehby GL, Murray JC (2008) Review on genetic variants and maternal smoking in the etiology of oral clefts and other birth defects. Birth Defects Res C 84:16–29CrossRefGoogle Scholar
  13. Skerjanc IS (1999) Cardiac and skeletal muscle development in P19 embryonal carcinoma cells. Trends Cardiovasc Med 9:139–143CrossRefPubMedGoogle Scholar
  14. Sullivan PM, Dervan LA, Reiger S, Buddhe S, Schwartz SM et al (2015) Risk of congenital heart defects in the offspring of smoking mothers: a population-based study. J Pediatr 166:978.e2–984.e2Google Scholar
  15. Volinia S, Calin GA, Liu C-G, Ambs S, Cimmino A, Petrocca F, Visone R, Iorio M, Roldo C, Ferracin M (2006) A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci USA 103:2257–2261CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  1. 1.Department of Cardiovascular SurgeryHenan Provincial People’s Hospital, People’s Hospital of Zhengzhou UniversityZhengzhouPeople’s Republic of China

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