Download PDF
Eur J Pediatr Surg 2020; 30(06): 524-528
DOI: 10.1055/s-0039-1700547
Original Article

CO2 Pneumoperitoneum Effects on Molecular Markers of Tumor Invasiveness in SH-SY5Y Neuroblastoma Cells

Angela Simona Montalto
1   Department of Human Pathology of Adult and Childhood “Gaetano Barresi,” University of Messina, Messina, Italy
,
Monica Currò
2   Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Messina, Italy
,
Tiziana Russo
1   Department of Human Pathology of Adult and Childhood “Gaetano Barresi,” University of Messina, Messina, Italy
,
Nadia Ferlazzo
2   Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Messina, Italy
,
Daniela Caccamo
2   Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Messina, Italy
,
Riccardo Ientile
2   Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Messina, Italy
,
Carmelo Romeo
1   Department of Human Pathology of Adult and Childhood “Gaetano Barresi,” University of Messina, Messina, Italy
,
Pietro Impellizzeri
1   Department of Human Pathology of Adult and Childhood “Gaetano Barresi,” University of Messina, Messina, Italy
› Author Affiliations
› Further Information
    Permissions and Reprints

Abstract

Introduction CO2 pneumoperitoneum can influence the biological behavior of neuroblastoma (NB). Angiogenesis and genetic features are responsible for malignant phenotype of this tumor. We examined the CO2 effects on N-Myc, vascular endothelial growth factor (VEGF), and matrix metalloproteinase-2 (MMP-2) expression as critical biomarkers of tumor invasiveness, in NB cells without N-Myc amplification.

Materials and Methods SH-SY5Y cells were exposed to CO2 (100%) at 15 mm Hg pressure for 4 hours and then moved to normal condition for 24 hours. Control cells were incubated with 5% CO2 for the same time. In control and CO2-exposed cells, the messenger ribonucleic acid (mRNA) levels of hypoxia-inducible factor (HIF)-1α, HIF-2α, VEGF-A, and MMP-2 were quantified by real-time polymerase chain reaction. N-Myc expression was evaluated by Western blot analysis.

Results The exposure to 15 mm Hg CO2 (100%) for 4 hours induced an increase in HIF-1α, but not in HIF-2α, mRNA levels. No differences were observed in N-Myc expression between exposed and control cells at each incubation time. Similarly, no significant differences were found for VEGF-A and MMP-2 transcript levels. In CO2 exposed cells, we observed only a slight increase in both VEGF-A and MMP-2 mRNA levels after 4 and 24 hours in comparison to controls.

Conclusion In our study, the hypoxic environment induced by CO2 exposure does not affect the expression of critical biomarkers of NB aggressiveness, such as N-Myc, VEGF, and MMP-2, in human SH-SY5Y NB cells without N-Myc amplification. These data suggest that CO2 pneumoperitoneum might not adversely impact NB cell invasiveness; however, it is necessary to evaluate these effects in others in vitro and in vivo models.

Keywords

neuroblastoma cell - CO2 - pneumoperitoneum - angiogenesis - oncogene


Publication History

Received: 20 June 2018

Accepted: 17 September 2019

Article published online:
10 November 2019

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Maris JM. Recent advances in neuroblastoma. N Engl J Med 2010; 362 (23) 2202-2211
    CrossrefPubMedGoogle Scholar
  • 2 Luo Y-B, Cui X-C, Yang L, Zhang D, Wang JX. Advances in the surgical treatment of neuroblastoma. Chin Med J (Engl) 2018; 131 (19) 2332-2337
    CrossrefPubMedGoogle Scholar
  • 3 Metzelder M, Kuebler J, Shimotakahara A, Vieten G, von Wasielewski R, Ure BM. CO(2) pneumoperitoneum increases systemic but not local tumor spread after intraperitoneal murine neuroblastoma spillage in mice. Surg Endosc 2008; 22 (12) 2648-2653
    CrossrefPubMedGoogle Scholar
  • 4 Reismann M, Wehrmann F, Schukfeh N, Kuebler JF, Ure B, Glüer S. Carbon dioxide, hypoxia and low pH lead to overexpression of c-myc and HMGB-1 oncogenes in neuroblastoma cells. Eur J Pediatr Surg 2009; 19 (04) 224-227
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Majmundar AJ, Wong WJ, Simon MC. Hypoxia-inducible factors and the response to hypoxic stress. Mol Cell 2010; 40 (02) 294-309
    CrossrefPubMedGoogle Scholar
  • 6 Montalto AS, Currò M, Russo T. et al. In vitro CO2-induced ROS production impairs cell cycle in SH-SY5Y neuroblastoma cells. Pediatr Surg Int 2013; 29 (01) 51-59
    CrossrefPubMedGoogle Scholar
  • 7 Currò M, Montalto AS, Impellizzeri P. et al. CO(2) pneumoperitoneum induces in vitro hypoxic response culminating in apoptosis of human neuroblastoma cells. J Biol Regul Homeost Agents 2014; 28 (03) 497-506
    PubMedGoogle Scholar
  • 8 Poljaková J, Groh T, Gudino ZO. et al. Hypoxia-mediated histone acetylation and expression of N-myc transcription factor dictate aggressiveness of neuroblastoma cells. Oncol Rep 2014; 31 (04) 1928-1934
    CrossrefPubMedGoogle Scholar
  • 9 Krock BL, Skuli N, Simon MC. Hypoxia-induced angiogenesis: good and evil. Genes Cancer 2011; 2 (12) 1117-1133
    CrossrefPubMedGoogle Scholar
  • 10 Ziello JE, Jovin IS, Huang Y. Hypoxia-Inducible Factor (HIF)-1 regulatory pathway and its potential for therapeutic intervention in malignancy and ischemia. Yale J Biol Med 2007; 80 (02) 51-60
    PubMedGoogle Scholar
  • 11 Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell 2010; 141 (01) 52-67
    CrossrefPubMedGoogle Scholar
  • 12 Pattyn F, Speleman F, De Paepe A, Vandesompele J. RTPrimerDB: the real-time PCR primer and probe database. Nucleic Acids Res 2003; 31 (01) 122-123
    CrossrefPubMedGoogle Scholar
  • 13 Fuchs J, Schafbuch L, Ebinger M, Schäfer JF, Seitz G, Warmann SW. Minimally invasive surgery for pediatric tumors - current state of the art. Front Pediatr 2014; 2: 48
    CrossrefPubMedGoogle Scholar
  • 14 Trigg RM, Turner SD. ALK in neuroblastoma: biological and therapeutic implications. Cancers (Basel) 2018; 10 (04) 113
    CrossrefPubMedGoogle Scholar
  • 15 Boo YJ, Goedecke J, Muensterer OJ. Pediatric oncologic endosurgery. Int J Surg Oncol (N Y) 2017; 2 (07) e30
    CrossrefPubMedGoogle Scholar
  • 16 Schmidt AI, Reismann M, Kübler JF. et al. Exposure to carbon dioxide and helium reduces in vitro proliferation of pediatric tumor cells. Pediatr Surg Int 2006; 22 (01) 72-77
    CrossrefPubMedGoogle Scholar
  • 17 Reismann M, Lehmann D, Quandte M, Ure BM, Glüer S. Carbon dioxide does not affect the methylation status of prognostic important oncogenes Rassf1A and DCR2 in neuroblastoma cells. Pediatr Surg Int 2008; 24 (12) 1327-1330
    CrossrefPubMedGoogle Scholar
  • 18 Nilsson H, Jögi A, Beckman S, Harris AL, Poellinger L, Påhlman S. HIF-2alpha expression in human fetal paraganglia and neuroblastoma: relation to sympathetic differentiation, glucose deficiency, and hypoxia. Exp Cell Res 2005; 303 (02) 447-456
    CrossrefPubMedGoogle Scholar
  • 19 Noguera R, Fredlund E, Piqueras M. et al. HIF-1alpha and HIF-2alpha are differentially regulated in vivo in neuroblastoma: high HIF-1alpha correlates negatively to advanced clinical stage and tumor vascularization. Clin Cancer Res 2009; 15 (23) 7130-7136
    CrossrefPubMedGoogle Scholar
  • 20 Koh MY, Powis G. Passing the baton: the HIF switch. Trends Biochem Sci 2012; 37 (09) 364-372
    CrossrefPubMedGoogle Scholar
  • 21 Adhikary S, Eilers M. Transcriptional regulation and transformation by Myc proteins. Nat Rev Mol Cell Biol 2005; 6 (08) 635-645
    CrossrefPubMedGoogle Scholar
  • 22 Beltran H. The N-myc Oncogene: maximizing its targets, regulation, and therapeutic potential. Mol Cancer Res 2014; 12 (06) 815-822
    CrossrefPubMedGoogle Scholar
  • 23 Li D, Mei H, Qi M. et al. FOXD3 is a novel tumor suppressor that affects growth, invasion, metastasis and angiogenesis of neuroblastoma. Oncotarget 2013; 4 (11) 2021-2044
    CrossrefPubMedGoogle Scholar
  • 24 Kang J, Rychahou PG, Ishola TA, Mourot JM, Evers BM, Chung DH. N-myc is a novel regulator of PI3K-mediated VEGF expression in neuroblastoma. Oncogene 2008; 27 (28) 3999-4007
    CrossrefPubMedGoogle Scholar
  • 25 Chetty C, Lakka SS, Bhoopathi P, Rao JS. MMP-2 alters VEGF expression via alphaVbeta3 integrin-mediated PI3K/AKT signaling in A549 lung cancer cells. Int J Cancer 2010; 127 (05) 1081-1095
    CrossrefPubMedGoogle Scholar