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The importance of abrogation of G2-phase arrest in combined effect of TRAIL and ionizing radiation

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Řezáčová M., Vávrová J., Vokurková D., Tichý A., Knížek J., Psutka J.

Acta Biochimica Polonica

2005

vol. 52

issue 4

889-895

Background: In this work we studied the relationship between the enhanced expression of DR5 receptor and the effect of combination of TRAIL and ionizing radiation on cell cycle arrest and apoptosis induction in human leukemia cell line HL-60. Material and methods: DR5, APO2.7 and cell cycle were analyzed by flow cytometry. Proteins Bid and Mcl-1 were analyzed by Western-blotting. For clonogenic survival, colony assay on methylcellulose was used. Results: Ionizing radiation caused significantly enhanced positivity of DR5 receptors 24 h after irradiation with high doses (6 and 8 Gy). An increase of DR5 receptor positivity after a dose of 2 Gy was not statistically significant and application of TRAIL 48 h after irradiation did not increase the apoptosis induction. However, a decrease of radiation-induced G2 phase arrest and an increase of apoptosis were observed when TRAIL was applied 16 h before irradiation with the dose of 2 Gy. Incubation with 6 µg/l TRAIL for 16 h reduced D0 value from 2.9 Gy to 1.5 Gy. The induction of apoptosis by TRAIL was accompanied by Bid cleavage and a decrease of antiapoptotic Mcl-1 16 h after incubation with TRAIL. Conclusion: TRAIL in concentration of 6 µg/l applied 16 h before irradiation by the dose of 1.5 Gy caused the death of 63% of clonogenic tumor cells, similarly as the dose of 2.9 Gy alone, which is in good correlation with the enhanced apoptosis induction.

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1 Acta Biochimica Polonica

1 2005

1 Knížek J.

1 Psutka J.

1 Tichý A.

1 Vokurková D.

1 Vávrová J.

1 Řezáčová M.

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The importance of abrogation of G2-phase arrest in combined effect of TRAIL and ionizing radiation