PURPOSE: To quantify the effects of irradiation on the expression levels of a differentially expressed gene, RTP801, in HeLa cells. MATERIALS AND METHODS: Total RNA was isolated from irradiated and non-irradiated HeLa cells. A subtraction library was constructed, from which 88 random clones were screened. The expression patterns of one clone, detected by reverse Northern blotting, were quantified by real time RT-PCR, using CYBR green I dye. RESULTS: RTP801, a hypoxia-inducible factor-I-responsive gene, was identified as a differentially expressed gene in HeLa cells exposed to X-ray. Real time RT-PCR showed that the mRNA levels of RTP801 were greatly diminished by radiation. CONCLUSIONS: These results suggest that down-regulation of hypoxia-inducible factor-I-responsive genes, such as RTP801, in irradiated HeLa cells may result in reductions in the radiotherapy resistance of tumor cells.
PURPOSE: To quantify the effects of irradiation on the expression levels of a differentially expressed gene, RTP801, in HeLa cells. MATERIALS AND METHODS: Total RNA was isolated from irradiated and non-irradiated HeLa cells. A subtraction library was constructed, from which 88 random clones were screened. The expression patterns of one clone, detected by reverse Northern blotting, were quantified by real time RT-PCR, using CYBR green I dye. RESULTS: RTP801, a hypoxia-inducible factor-I-responsive gene, was identified as a differentially expressed gene in HeLa cells exposed to X-ray. Real time RT-PCR showed that the mRNA levels of RTP801 were greatly diminished by radiation. CONCLUSIONS: These results suggest that down-regulation of hypoxia-inducible factor-I-responsive genes, such as RTP801, in irradiated HeLa cells may result in reductions in the radiotherapy resistance of tumor cells.
Entities:
Keywords:
HeLa cells; RTP801; Radiation; Real time PCR
Authors: Claudia E Rübe; Frans van Valen; Falk Wilfert; Jan Palm; Andreas Schuck; Normann Willich; Winfried Winkelmann; Heribert Jürgens; Christian Rübe Journal: Int J Radiat Oncol Biol Phys Date: 2003-08-01 Impact factor: 7.038