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Silencing of FABP3 Inhibits Proliferation and Promotes Apoptosis in Embryonic Carcinoma Cells

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Abstract

Fatty acid–binding protein 3 (FABP3) facilitates the movement of fatty acids in cardiac muscle. Previously, we reported that FABP3 is highly upregulated in the myocardium of ventricular septal defect patients and overexpression of FABP3 inhibited proliferation and promoted apoptosis in embryonic carcinoma cells (P19 cells). In this study, we aimed to investigate the effect of FABP3 gene silencing on P19 cell differentiation, proliferation and apoptosis. We used RNA interference and a lentiviral-based vector system to create a stable FABP3-silenced P19 cell line; knockdown of FABP3 was confirmed by quantitative real-time PCR. Expression analysis of specific differentiation marker genes using quantitative real-time PCR and observation of morphological changes using an inverted microscope revealed that knockdown of FABP3 did not significantly affect the differentiation of P19 cells into cardiomyocytes. CCK-8 proliferation assays and cell cycle analysis demonstrated that FABP3 gene silencing significantly inhibited P19 cell proliferation. Furthermore, Annexin V-FITC/propidium iodide staining and the caspase-3 activity assay revealed that FABP3 gene silencing significantly promoted serum starvation–induced apoptosis in P19 cells. In agreement with our previous research, these results demonstrate that FABP3 may play an important role during embryonic heart development, and that either overexpression or silencing of FABP3 will lead to an imbalance between proliferation and apoptosis, which may result in embryonic cardiac malformations.

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References

  1. Olson, E. N. (2006). Gene regulatory networks in the evolution and development of the heart. Science, 313, 1922–1927.

    Article  PubMed  CAS  Google Scholar 

  2. Hoffman, J. I., & Kaplan, S. (2002). The incidence of congenital heart disease. Journal of the American College of Cardiology, 39, 1890–1900.

    Article  PubMed  Google Scholar 

  3. Garg, V. (2006). Insights into the genetic basis of congenital heart disease. Cellular and Molecular Life Sciences, 63, 1141–1148.

    Article  PubMed  CAS  Google Scholar 

  4. Sands, A. J., Casey, F. A., Craig, B. G., Dornan, J. C., Rogers, J., & Mulholland, H. C. (1999). Incidence and risk factors for ventricular septal defect in “low risk” neonates. Archives of Disease in Childhood. Fetal and Neonatal Edition, 81, F61–F63.

    Article  PubMed  CAS  Google Scholar 

  5. Kaynak, B., von Heydebreck, A., Mebus, S., Seelow, D., Hennig, S., Vogel, J., et al. (2003). Genome-wide array analysis of normal and malformed human hearts. Circulation, 107, 2467–2474.

    Article  PubMed  Google Scholar 

  6. Besnard, P., Niot, I., Poirier, H., Clement, L., & Bernard, A. (2002). New insights into the fatty acid-binding protein (FABP) family in the small intestine. Molecular and Cellular Biochemistry, 239, 139–147.

    Article  PubMed  CAS  Google Scholar 

  7. Qian, Q., Kuo, L., Yu, Y. T., & Rottman, J. N. (1999). A concise promoter region of the heart fatty acid-binding protein gene dictates tissue-appropriate expression. Circulation Research, 84, 276–289.

    Article  PubMed  CAS  Google Scholar 

  8. Zhang, H., Zhou, L., Yang, R., Sheng, Y., Sun, W., Kong, X., et al. (2006). Identification of differentially expressed genes in human heart with ventricular septal defect using suppression subtractive hybridization. Biochemical and Biophysical Research Communications, 342, 135–144.

    Article  PubMed  CAS  Google Scholar 

  9. Zhu, C., Hu, D. L., Liu, Y. Q., Zhang, Q. J., Chen, F. K., Kong, X. Q., et al. (2011). Fabp3 inhibits proliferation and promotes apoptosis of embryonic myocardial cells. Cell Biochemistry and Biophysics, 60, 259–266.

    Article  PubMed  CAS  Google Scholar 

  10. van der Heyden, M. A., van Kempen, M. J., Tsuji, Y., Rook, M. B., Jongsma, H. J., & Opthof, T. (2003). P19 embryonal carcinoma cells: A suitable model system for cardiac electrophysiological differentiation at the molecular and functional level. Cardiovascular Research, 58, 410–422.

    Article  PubMed  Google Scholar 

  11. Grepin, C., Nemer, G., & Nemer, M. (1997). Enhanced cardiogenesis in embryonic stem cells overexpressing the GATA-4 transcription factor. Development, 124, 2387–2395.

    PubMed  CAS  Google Scholar 

  12. Chen, C., & Okayama, H. (1987). High-efficiency transformation of mammalian cells by plasmid DNA. Molecular and Cellular Biology, 7, 2745–2752.

    PubMed  CAS  Google Scholar 

  13. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) Method. Methods, 25, 402–408.

    Article  PubMed  CAS  Google Scholar 

  14. Vermes, I., Haanen, C., Steffens-Nakken, H., & Reutelingsperger, C. (1995). A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. Journal of Immunological Methods, 184, 39–51.

    Article  PubMed  CAS  Google Scholar 

  15. Koike, M., Sakaki, S., Amano, Y., & Kurosawa, H. (2007). Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies. Journal of Bioscience and Bioengineering, 104, 294–299.

    Article  PubMed  CAS  Google Scholar 

  16. Lints, T. J., Parsons, L. M., Hartley, L., Lyons, I., & Harvey, R. P. (1993). Nkx-2.5: A novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants. Development, 119, 419–431.

    PubMed  CAS  Google Scholar 

  17. Bruneau, B. G., Nemer, G., Schmitt, J. P., Charron, F., Robitaille, L., Caron, S., et al. (2001). A murine model of Holt-Oram syndrome defines roles of the T-box transcription factor Tbx5 in cardiogenesis and disease. Cell, 106, 709–721.

    Article  PubMed  CAS  Google Scholar 

  18. Watt, A. J., Battle, M. A., Li, J., & Duncan, S. A. (2004). GATA4 is essential for formation of the proepicardium and regulates cardiogenesis. Proceedings of the National Academy of Sciences of the United States of America, 101, 12573–12578.

    Article  PubMed  CAS  Google Scholar 

  19. Tang, M. K., Kindler, P. M., Cai, D. Q., Chow, P. H., Li, M., & Lee, K. K. (2004). Heart-type fatty acid binding proteins are upregulated during terminal differentiation of mouse cardiomyocytes, as revealed by proteomic analysis. Cell and Tissue Research, 316, 339–347.

    Article  PubMed  CAS  Google Scholar 

  20. Aagaard, L., & Rossi, J. J. (2007). RNAi therapeutics: Principles, prospects and challenges. Advanced Drug Delivery Reviews, 59, 75–86.

    Article  PubMed  CAS  Google Scholar 

  21. Dykxhoorn, D. M., Novina, C. D., & Sharp, P. A. (2003). Killing the messenger: Short RNAs that silence gene expression. Nature Reviews Molecular Cell Biology, 4, 457–467.

    Article  PubMed  CAS  Google Scholar 

  22. Abbas-Terki, T., Blanco-Bose, W., Deglon, N., Pralong, W., & Aebischer, P. (2002). Lentiviral-mediated RNA interference. Human Gene Therapy, 13, 2197–2201.

    Article  PubMed  CAS  Google Scholar 

  23. Brummelkamp, T. R., Bernards, R., & Agami, R. (2002). A system for stable expression of short interfering RNAs in mammalian cells. Science, 296, 550–553.

    Article  PubMed  CAS  Google Scholar 

  24. Zhu, C., Hu, D. L., Liu, Y. Q., Zhang, Q. J., Chen, F. K., Kong, X. Q., et al. (2011). Fabp3 inhibits proliferation and promotes apoptosis of embryonic myocardial cells. Cell Biochemistry and Biophysics, 60, 259–266.

    Article  PubMed  CAS  Google Scholar 

  25. Shen, Y. H., Song, G. X., Liu, Y. Q., Sun, W., Zhou, L. J., Liu, H. L., et al. (2012). Silencing of FABP3 promotes apoptosis and induces mitochondrion impairment in embryonic carcinoma cells. Journal of Bioenergetics and Biomembranes, 44, 317–323.

    Article  PubMed  CAS  Google Scholar 

  26. Song, G. X., Shen, Y. H., Liu, Y. Q., Sun, W., Miao, L. P., Zhou, L. J., Liu, H. L., Yang, R., Kong, X. Q., Cao, K. J., Qian, L. M., & Sheng, Y. H. (2012). Overexpression of FABP3 promotes apoptosis through inducing mitochondrial impairment in embryonic cancer cells. Journal of Cellular Biochemistry, Jul 2. doi: 10.1002/jcb.24243. [Epub ahead of print].

  27. Srivastava, D. (2006). Genetic regulation of cardiogenesis and congenital heart disease. Annual Review of Pathology: Mechanisms of Disease, 1, 199–213.

    Article  CAS  Google Scholar 

  28. Fiorina, P., Corradi, D., Pinelli, S., Maestri, R., Lagrasta, C., Buscaglia, M., et al. (2004). Apoptotic/mytogenic pathways during human heart development. International Journal of Cardiology, 96, 409–417.

    Article  PubMed  Google Scholar 

  29. Levy, M., Maurey, C., Celermajer, D. S., Vouhe, P. R., Danel, C., Bonnet, D., et al. (2007). Impaired apoptosis of pulmonary endothelial cells is associated with intimal proliferation and irreversibility of pulmonary hypertension in congenital heart disease. Journal of the American College of Cardiology, 49, 803–810.

    Article  PubMed  CAS  Google Scholar 

  30. Gittenberger-de, G. A., Bartelings, M. M., Deruiter, M. C., & Poelmann, R. E. (2005). Basics of cardiac development for the understanding of congenital heart malformations. Pediatric Research, 57, 169–176.

    Article  Google Scholar 

  31. Hoye, A. T., Davoren, J. E., Wipf, P., Fink, M. P., & Kagan, V. E. (2008). Targeting mitochondria. Accounts of Chemical Research, 41, 87–97.

    Article  PubMed  CAS  Google Scholar 

  32. McBurney, M. W., Jones-Villeneuve, E. M., Edwards, M. K., & Anderson, P. J. (1982). Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature, 299, 165–167.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 81070138), the Natural Science Foundation of Jiangsu Province, China (No. BK2010582) and the Talent Foundation of Jiangsu Province, China (WSN-020).

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Authors and Affiliations

  1. State Key Laboratory of Reproductive Medicine, Department of MICU, Nan**g Maternity and Child Health Care Hospital Affiliated to Nan**g Medical University, Nan**g, 210029, China

    Yahui Shen & Yaoqiu Liu

  2. Department of Cardiology, The First Affiliated Hospital of Nan**g Medical University, Nan**g, 210029, China

    Guixian Song, Lijuan Zhou, Hailang Liu, **angqing Kong, Yanhui Sheng, Kejiang Cao & Lingmei Qian

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  1. Yahui Shen
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  2. Guixian Song
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  3. Yaoqiu Liu
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  6. **angqing Kong
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Correspondence to Lingmei Qian.

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Yahui Shen, Guixian Song contributed equally to this work.

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Shen, Y., Song, G., Liu, Y. et al. Silencing of FABP3 Inhibits Proliferation and Promotes Apoptosis in Embryonic Carcinoma Cells. Cell Biochem Biophys 66, 139–146 (2013). https://doi.org/10.1007/s12013-012-9462-y

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