Eur J Pediatr Surg 2009; 19(4): 224-227
DOI: 10.1055/s-0029-1202778
Original Article

© Georg Thieme Verlag KG Stuttgart · New York

Carbon dioxide, Hypoxia and Low pH Lead to Overexpression of c-myc and HMGB-1 Oncogenes in Neuroblastoma Cells

M. Reismann1 , F. Wehrmann1 , N. Schukfeh1 , J. F. Kuebler1 , B. Ure1 , S. Glüer1
  • 1Department of Pediatric Surgery, Medical School Hannover, Hannover, Germany
Further Information

Publication History

received December 8, 2008

accepted after revision January 6, 2009

Publication Date:
22 April 2009 (online)

Abstract

Purpose: It has been reported that CO2 gas, used to establish a pneumoperitoneum during laparoscopy, affects the behavior of tumor cells. The proto-oncogenes c-myc and HMGB-1 mediate aggressive behavior of neuroblastomas. We studied whether exposure to CO2, hypoxia or acidosis affects the expression of c-myc and HMGB-1 in neuroblastoma cells.

Methods: SH-SY5Y cells were incubated with 100% CO2, 95% helium/5% CO2 or pH 6.2 for 2 h. The expression of c-myc and HMGB-1 was measured by Western blot test immediately, 3 h and 6 h after incubation. Additionally, we measured apoptosis after incubation using fluorometric measurements of caspase 3 and 7 activity.

Results: C-myc (160±26%, p=0.007 and 138±16% vs. control, p=0.04) and HMGB-1 proteins (140±13% and 136±11%, both p=0.037) were found to be significantly upregulated 6 h after incubation with CO2 and helium. There was early upregulation of both oncogenes 3 h after CO2 incubation (251±79%, p=0.04 and 292±136, p=0.037). Correspondingly, pH 6.2 led to significant overexpression. Levels of apoptosis were reduced.

Conclusions: Exposures mimicking conditions of CO2 pneumoperitoneum lead to significant overexpression of c-myc and HMGB-1 in neuroblastoma cells with decreased apoptosis. These results point to a negative influence and potentially increased malignancy of tumor cells.

References

  • 1 Bártová E, Kozubek S, Kozubek M, Jirsová P, Lukásová E, Skalníková M, Cafourková A. Koutná I. Nuclear topography oft the c-myc gene in human leukemic cells.  Gene. 2000;  244 1-11
  • 2 Cirocco WC, Schwartzman A, Golub RW. Abdominal wall recurrence after laparoscopic colectomy for colon cancer.  Surgery. 1994;  116 842-846
  • 3 Conzen SD, Gottlob K, Kandel ES, Khanduri P, Wagner AJ, O’Leary M, Hay N. Induction of cell cycle progression and acceleration of apoptosis are two separable functions of c-myc: transrepression correlates with acceleration of apoptosis.  Mol Cell Biol. 2000;  20 6008-6018
  • 4 Lagausie P de, Berrebi D, Michon J, Philippe-Chomette P, El Ghoneimi A, Garel C, Brisse H, Peuchmaur M, Aigrain Y. Laparoscopic adrenal surgery for neuroblastomas in children.  J Urol. 2003;  170 932-935
  • 5 Eberhart CG, Kratz J, Wang Y, Summers K, Stearns D, Cohen K, Dang CV, Burger PC. Histopathological and molecular prognostic markers in medulloblastoma: c-myc, N-myc, TrkC, an anaplasia.  J Neuropathol Exp Neurol. 2004;  63 441-449
  • 6 Gleeson NC, Nicosia SV, Mark JE, Hoffman MS, Cavanagh D. Abdominal wall metastases from ovarian cancer after laparoscopy.  Am J Obstet Gynecol. 1993;  169 522-523
  • 7 Holmquist L, Lofstedt T, Pahlman. Effect of hypoxia on the phenoptype: the neuroblastoma and breast cancer models.  Adv Exp Med Biol. 2006;  587 179-193
  • 8 Iwanaka T, Arai M, Yamamoto H, Fukuzawa M, Kubota A, Kouchi K, Nio M, Satomi A, Sasaki F, Yoneda A, Ohhama Y, Takehara H, Morikawa Y, Miyano T. No incidence of port-site recurrence after endosurgical procedure for pediatric malignancies.  Pediatr Surg Int. 2003;  19 ((3)) 200-203
  • 9 Iwanaka T, Arya G, Ziegler MM. Mechanism and prevention of port-site tumor recurrence after laparoscopy in a murine model.  J Ped Surg. 1998;  33 457-461
  • 10 Kos M, Kuebler JF, Jesch NK, Vieten G, Bax NM, Zee DC van der, Busche R, Ure BM. Carbon dioxide differentially affects the cytokine release of macrophage subpopulations exclusively via alteration of extracellular pH.  Surg Endosc. 2006;  20 570-576
  • 11 Kuhry E, Schwenk W, Gaupset R, Romild U, Bonjer J. Long-term outcome of laparoscopic surgery for colorectal cancer: a Cochrane systematic review of randomised controlled trials.  Cancer Treat Rev. 2008;  34 498-504
  • 12 Larcher JC, Vayssière JL, Lossouarn L, Gros F, Croizat B. Regulation of c- and N-myc expression during induced differentiation of murine neuroblastoma cells.  Onkogene. 1991;  6 633-638
  • 13 Liu J, Narasimhan P, Lee YS, Song YS, Endo H, Yu F, Chan PH. Mild hypoxia promotes survival and proliferation of SOD2-deficient astrocyts via c-myc activation.  J Neuroscience. 2006;  26 4329-4337
  • 14 Metzelder ML, Kuebler JF, Shimotakahara A, Glueer S, Grigull L, Ure BM. Role of diagnostic and ablative minimally invasive surgery for pediatric malignancies.  Cancer. 2007;  109 2343-2348
  • 15 Metzelder ML, Kuebler JF, Shimotakahara A, Wasilewski R, Vieten G, Ure BM. CO2 pneumoperitoneum increases systemic but not local tumor spread after intraperitoneal murine neuroblastoma spillage in mice.  Surg Endosc. 2008;  , [in press]
  • 16 Metzelder ML, Ure BM. Port-site metastasis after laparoscopic biopsy of post transplant Burkitt lymphoma in a child.  Eur J Ped Surg. 2008;  , [in press]
  • 17 Mossafa H, Damotte D, Jenabian A, Delarue R, Vincenneau A, Amouroux I, Jeandel R, Khoury E, Martelli JM, Samson T, Tapia S, Flandrin G. Troussard X. Non-Hodgkin's lymphomas with Burkitt-like cells are associated with c-myc amplification and poor prognosis.  Leuk Lymphoma. 2006;  47 1885-1893
  • 18 Muntz HG, Goff BA, Madsen BL, Yon JL. Port site recurrence after laparoscopic surgery for endometrial carcinoma.  Obstet Gynecol. 1999;  93 807-809
  • 19 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 1327-1330
  • 20 Rothermund K, Rogulski K, Fernandes E, Whiting A, Sedivy J, Pu L, Prochownik EV. C-myc-independent restoration of multiple phenotypes by two c-myc target genes with overlapping functions.  Cancer Res. 2005;  65 2097-2107
  • 21 Sapan CV, Lundblad R L, Price NC. Colometric protein assay techniques.  Biotechnol Appl Biochem. 1999;  29 99-108
  • 22 Schmidt AI, Reismann M, Kübler JF, Vieten G, Bangen C, Shimotakahara A, Glüer S, Nustede R, Ure BM. Exposure to carbon dioxide and helium reduces in vitro proliferation of pediatric tumor cells.  Pediatr Surg Int. 2006;  22 72-77
  • 23 Schuller HM, Plummer III HK, Jull BA. Receptor-mediated effects of nicotine and its nitrosated derivative NNK on pulmonary neuroendocrine cells.  Anat Rec Part A Discov Mol Cell Eurl Biol. 2003;  270 51-58
  • 24 Scotting PJ, Walker DA, Perilongo G. Childhood solid tumors: a developmental disorder.  Nat Rev Cancer. 2005;  5 481
  • 25 Stennicke HR, Salnvesen GS. Biochemical characteristics of caspase-3, -6, -7, and -8.  J Biol Chem. 1997;  41 25719-25723
  • 26 Ure BM, Suempelmann R, Metzelder MM, Kuebler J. Physiologic responses to endoscopic surgery in children.  Sem Ped Surg. 2007;  16 217-223.
  • 27 Wildbrett P, Oh A, Naundorf D, Volk T, Jacobi CA. Impact of laparoscopic gases on peritoneal microenvironment and essential parameters of cell function.  Surg Endosc. 2003;  17 78-82
  • 28 Wood LJ, Mukherjee M, Dolde CE, Xu Y, Maher JF, Bunton TE, Williams JB, Resar LM. HMG-I/Y, a new c-myc target gene and potential oncogene.  Mol Cell Biol. 2000;  20 5490-5502

Correspondence

Dr. M. ReismannMD 

Department of Pediatric Surgery

Medical School Hannover

Carl-Neuberg-Str. 1

30625 Hannover

Germany

Phone: +49/0/511 532 92 60

Fax: +49/0/511 532 90 59

Email: reismann.marc@mh-hannover.de