title: Reoxygenation of human coronary smooth muscle cells suppresses HIF-1alpha gene expression and augments radiation-induced growth delay and apoptosis.
abstract: OBJECTIVE
  Catheter-based coronary brachytherapy with beta- and gamma-radiation is an evidence-based method to prevent restenosis after percutaneous transluminal coronary angioplasty (PTCA) and stent implantation, but the outcome may be subject to improvements. Physiological studies suggest that most of the target cells of brachytherapy in coronary arteries after PTCA are hypoxic. A lack of oxygen decreases the effect of low LET (linear energy transfer) irradiation. The authors assumed that reoxygenation of hypoxic human coronary smooth muscle cells (HCSMCs) improves the results of coronary brachytherapy. The expression of hypoxia-inducible factor 1alpha (HIF-1alpha) gene, and the rates of growth and apoptosis of hypoxic and reoxygenated HCSMCs after gamma-irradiation were therefore analyzed.
  
  
  METHODS
  An in vitro model of megacolonies of HCSMCs was developed. After exposure to chronic hypoxia the HCSMCs were irradiated with graded doses of 2, 4, 8, and 16 Gy using a (60)Co source either under hypoxia (pO(2) < 3 mmHg) or after reoxygenation (pO(2) approximately 150 mmHg). RT-PCR (reverse transcription-polymerase chain reaction) analysis was used to quantify HIF-1alpha gene expression and the growth of HCSMC megacolonies was measured serially. The oxygen enhancement ratio (OER) was calculated from the specific growth delay. Apoptosis of HCSMCs was quantified by counting cells with specific DNA strand breaks using the TUNEL assay.
  
  
  RESULTS
  HIF-1alpha gene expression was markedly suppressed in reoxygenated cells versus hypoxic cells 30 min after gamma-irradiation at all radiation doses (158 +/- 46% vs. 1,675 +/- 1,211%; p < 0.01). Apoptosis was markedly increased in reoxygenated HCSMCs. The OER was 1.8 (95% CI [confidence interval] 1.3-2.4). Therefore, reoxygenated HCSMCs require 44% less radiation dose to achieve the equivalent biological radiation effect compared to hypoxic HCSMCs.
  
  
  CONCLUSIONS
  Reoxygenation of coronary smooth muscle cells should be considered an option to increase efficacy of coronary brachytherapy. This could be used to reduce radiation dose and associated late side effects.
authors.affiliation: Department of Cardiology, University Clinic of Freiburg, Germany. grumann@med1.ukl.uni-freiburg.de
authors.forename: Thorsten
authors.initials: T
authors.lastname: Grumann
authors.forename: Amina
authors.initials: A
authors.lastname: Arab
authors.forename: Christoph
authors.initials: C
authors.lastname: Bode
authors.forename: Roland
authors.initials: R
authors.lastname: Guttenberger
authors.forename: Christoph
authors.initials: C
authors.lastname: Hehrlein
mesh_terms: D015906:Angioplasty, Balloon, Coronary; D017209:Apoptosis; D001918:Brachytherapy; D015687:Cell Hypoxia; D002478:Cells, Cultured; D023903:Coronary Restenosis / Q000517:prevention & control*; D003331:Coronary Vessels / Q000378:metabolism / Q000528:radiation effects*; D003627:Data Interpretation, Statistical; D015870:Gene Expression; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D018499:Linear Energy Transfer; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism / Q000528:radiation effects*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000528:radiation effects*; D010100:Oxygen / Q000378:metabolism*; D011879:Radiotherapy Dosage; D015607:Stents
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010100:Oxygen