Skip to main content

Advertisement

SpringerLink
Log in

The Effect of Cigarette Smoke Extract on Trophoblast Cell Viability and Migration: The Role of Adrenomedullin

  • Original Articles
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

We tested the hypothesis that cigarette smoke extract (CSE) leads to differences in expression of genes involved in angiogenesis and affects cell viability and migration in a first-trimester cytotrophoblast cell line (HTR-8/SVneo). HTR-8/SVneo cells were treated with 1% CSE, and gene expression for adrenomedullin (ADM), placental growth factor (PIGF), soluble fms-like tyrosine kinase I (sFLT-1), and vascular endothelia growth factor (VEGF) and protein content for ADM, PIGF, and sFlt-1 determined. A cell viability assay and a cell migration scratch assay were utilized following treatment with CSE with and without ADM inhibitor. Adrenomedullin, PIGF, and VEGF gene transcripts were significantly upregulated by 1% CSE treatment compared with unstimulated cells or cells treated with nicotine alone. Neither 1% CSE nor nicotine treatment alone affected sFlt-1 gene expression. There was a significant increase in secreted ADM protein from cells treated with 1% CSE detected by enzyme-linked immuno-sorbant assay, though no differences in PIGF or sFIt-1 production were seen. Treatment with 1 % CSE increased cell viability and cell migration compared with unstimulated cells and was inhibited by co-treatment with ADM inhibitor. Treatment of a first-trimester trophoblast cell line with CSE increases cell viability and cell migration that are reversed by co-treatment with ADM inhibitor, suggesting that ADM at least partially mediates cell growth and viability following CSE treatment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Report of the National High Blood Pressure Education Program Working Group on High Blood Pressure in Pregnancy. Am J Obstet Gynecol. 2000;183(1):S1–S22.

  2. Sibai B, Dekker G, Kupferminc M. Pre-eclampsia. Lancet. 2005;365(9461):785–799.

    Article  Google Scholar 

  3. Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science. 2005;308:1592–1594.

    Article  CAS  Google Scholar 

  4. Arroyo JA, Winn VD. Vasculogenesis and angiogenesis in the IUGR placenta. Semin Perinatol. 2008;32(3):172–177.

    Article  Google Scholar 

  5. Shibuya M. Structure and function of VEGF/VEGF-receptor system involved in angiogenesis. Cell Struct Funct. 2001;26(1):25–35.

    Article  CAS  Google Scholar 

  6. Levine RJ, Maynard SE, Qian C, et al. Circulating angiogenic factors and the risk of preeclampsia. N Engl J Med. 2004;350(7):672–683.

    Article  CAS  Google Scholar 

  7. Gibbons C, Dackor R, Dunworth W, Fritz-Six K, Caron KM. Receptor activity-modifying proteins: RAMPing up adrenome-dullin signaling. Mol Endocrinol. 2007;21(4):783–796.

    Article  CAS  Google Scholar 

  8. Caron KM, Smithies O. Multiple roles of adrenomedullin revealed by animal models. Microsc Res Tech. 2002;57(1):55–59.

    Article  CAS  Google Scholar 

  9. Gratton RJ, Gluszynski M, Mazzuca DM, Nygard K, Han VK. Adrenomedullin messenger ribonucleic acid expression in the placentae of normal and preeclamptic pregnancies. J Clin Endocrinol Metab. 2003;88(12):6048–6055.

    Article  CAS  Google Scholar 

  10. Beltowski J, Jamroz A. Adrenomedullin—what do we know 10 years since its discovery? Pol J Pharmacol. 2004;56(1):5–27.

    CAS  PubMed  Google Scholar 

  11. Castles A, Adams EK, Melvin CL, Kelsch C, Boulton ML. Effects of smoking during pregnancy. Five meta-analyses. Am J Prev Med. 1999;16(3):208–215.

    Article  CAS  Google Scholar 

  12. Mehendale R, Hibbard J, Fazleabas A, Leach R. Placental angio-genesis markers sFlt-1 and P1GF: response to cigarette smoke. Am J Obstet Gynecol. 2007;197:363. e1–e5.

    Article  Google Scholar 

  13. Graham CH, Hawley TS, Hawley RG, et al. Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 1993;206(2):204–211.

    Article  CAS  Google Scholar 

  14. Munaut C, Lorquet S, Pequeux C, et al. Hypoxia is responsible for soluble vascular endothelial growth factor receptor-1 (VEGFR-1) but not for soluble endoglin induction in villous trophoblast. Hum Reprod. 2008;23(6):1407–1415.

    Article  CAS  Google Scholar 

  15. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2007;2(2):329–333.

    Article  CAS  Google Scholar 

  16. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29(9):e45.

    Article  CAS  Google Scholar 

  17. Mazzocchi G, Albertin G, Andreis PG, et al. Distribution, functional role, and signaling mechanism of adrenomedullin receptors in the rat adrenal gland. Peptides. 1999;20:1479–1487.

    Article  CAS  Google Scholar 

  18. Kim H, Liu X, Kobayashi T, et al. Reversible cigarette smoke extract-induced DNA damage in human lung fibroblasts. Am J Respir Cell Mol Biol. 2004;31(5):483–490.

    Article  CAS  Google Scholar 

  19. Shapiro SD. Smoke gets in your cells. Am J Respir Cell Mol Biol. 2004;31(5):481–482.

    Article  CAS  Google Scholar 

  20. England L, Zhang J. Smoking and risk of preeclampsia: a systematic review. Front Biosci. 2007;12:2471–2483.

    Article  CAS  Google Scholar 

  21. Conde-Agudelo A, Althabe F, Belizan JM, Kafury-Goeta AC. Cigarette smoking during pregnancy and risk of preeclampsia: a systematic review. Am J Obstet Gynecol. 1999;181(4):1026–1035.

    Article  CAS  Google Scholar 

  22. Wikstrom AK, Stephansson O, Cnattingius S. Tobacco use during pregnancy and preeclampsia risk: effects of cigarette smoking and snuff. Hypertension. 2010;55(5):1254–1259.

    Article  Google Scholar 

  23. Jeyabalan A, Powers RW, Durica AR, Harger GF, Roberts JM, Ness RB. Cigarette smoke exposure and angiogenic factors in pregnancy and preeclampsia. Am J Hypertens. 2008;21(8):943–947.

    Article  CAS  Google Scholar 

  24. Belgore FM, Lip GY, Blann AD. Vascular endothelial growth factor and its receptor, Flt-1, in smokers and non-smokers. Br J Biomed Sci. 2000;57(3):207–213.

    CAS  PubMed  Google Scholar 

  25. Caron KM, Smithies O. Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene. Proc Natl Acad Sci U S A. 2001;98(2):615–619.

    Article  CAS  Google Scholar 

  26. Li M, Yee D, Magnuson TR, Smithies O, Caron KM. Reduced maternal expression of adrenomedullin disrupts fertility, placentation, and fetal growth in mice. J Clin Invest. 2006;116(10):2653–2662.

    Article  CAS  Google Scholar 

  27. Lenhart P, Li M, Schwerbrock N, Fritz-Six K, Kadmiel M, Caron K. Fetal loss of adrenomedullin causes maternal vascular pathology typical of preeclampsia. Abstract, 44th Annual Meeting of the Society for the Study of Reproduction, Portland, OR August 3, 2011.

  28. Zhang X, Green KE, Yallampalli C, Dong YL. Adrenomedullin enhances invasion by trophoblast cell lines. Biol Reprod. 2005;73(4):619–626.

    Article  CAS  Google Scholar 

  29. Senna AA, Zedan M, el-Salam GE, el-Mashad AI. Study of plasma adrenomedullin level in normal pregnancy and preclampsia. Medscape J Med. 2008;10(2):29.

    PubMed  PubMed Central  Google Scholar 

  30. Dikensoy E, Balat O, Pence S, Balat A, Cekmen M, Yurekli M. The changes of plasma malondialdehyde, nitric oxide, and adrenomedullin levels in patients with preeclampsia. Hypertens Pregnancy. 2009;28(4):383–389.

    Article  CAS  Google Scholar 

  31. Alonso-Martinez JL, Anniccherico-Sanchez FJ, Urbieta-Echezarreta M, Garcia-Sanchotena JL, Olaz-Preciado F. Hemodynamic effects of chronic smoking in liver cirrhosis: a role for adrenomedullin. Eur J Gastroenterol Hepatol. 2010;22(5):513–518.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Duke University School of Medicine, DUMC Box 3967, Durham, NC, 27710, USA

    Timothy R. Beiswenger MD, Liping Feng MD, Haywood L. Brown MD, R. Phillips Heine MD, Amy P. Murtha MD & Chad A. Grotegut MD

Authors
  1. Timothy R. Beiswenger MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liping Feng MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Haywood L. Brown MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. Phillips Heine MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Amy P. Murtha MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chad A. Grotegut MD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chad A. Grotegut MD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Beiswenger, T.R., Feng, L., Brown, H.L. et al. The Effect of Cigarette Smoke Extract on Trophoblast Cell Viability and Migration: The Role of Adrenomedullin. Reprod. Sci. 19, 526–533 (2012). https://doi.org/10.1177/1933719111426600

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1177/1933719111426600

Keywords

Search

Navigation