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Effects of Hypobaric Conditions on Apoptosis Signalling Pathways in HeLa Cells

  • Arican, Gul Ozcan (Department of Biology, Faculty of Science, Istanbul University) ;
  • Khalilia, Walid (Programme of Radiobiology, Institute of Science, Istanbul University) ;
  • Serbes, Ugur (Programme of Radiobiology, Institute of Science, Istanbul University) ;
  • Akman, Gizem (Department of Biology, Faculty of Science, Istanbul University) ;
  • Cetin, Idil (Department of Biology, Faculty of Science, Istanbul University) ;
  • Arican, Ercan (Department of Molecular Biology and Genetics, Faculty of Science, Istanbul University)
  • Published : 2014.06.30

Abstract

Nowadays increasing effectiveness in cancer therapy and investigation of formation of new strategies that enhance antiproliferative activity against target organs has become a subject of interest. Although the molecular mechanisms of apoptosis can not be fully explained, it is known that cell suicide program existing in their memory genetically is activated by pathophysiological conditions and events such as oxidative stress. Low pressure (hypobaric) conditions that create hypoxia promote apoptosis by inhibiting cell cycling. In this study, determination of the effects of fractional hypobaric applications at different times on HeLa cells at cellular and molecular levels were targeted. Experiments were carried out under hypobaric conditions (35.2 kPa) in a specially designed hypobaric cabin including 2% $O_2$ and 98% N. Application of fractional hypobaric conditions was repeated two times for 3 hours with an interval of 24 hours. At the end of the implementation period cells were allowed to incubate for 24 hours for activation of repair mechanisms. Cell kinetic parameters such as growth rate (MTT) and apoptotic index were used in determination of the effect of hypobaric conditions on HeLa cells. Also in our study expression levels of the Bcl-2 gene family that have regulatory roles in apoptosis were determined by the RT-PCR technique to evaluate molecular mechanisms. The results showed that antiproliferative effect of hypobaric conditions on HeLa cells started three hours from the time of application and increased depending on the period of exposure. While there was a significant decrease in growth rate values, there was a significant increase in apoptotic index values (p<0.01). Also molecular studies showed that hypobaric conditions caused a significant increase in expression level of proapoptotic gene Bax and significant decrease in antiapoptotic Bfl-1. Consequently fractional application of hypobaric conditions on HeLa cell cultures increased both antiproliferative and apoptotic effects and these effects were triggered by the Bax gene.

Acknowledgement

Supported by : University of Istanbul

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