Unknown,Transcriptomics,Genomics,Proteomics

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Genome-wide gene expression analysis of HL60 cells and its alpha particle-resistant derivative RA11, gamma resistant-derivative RG8, and vincristine resistant-derivative RV+ at baseline and following 8Gy gamma IR and ~2Gy alpha IR

ABSTRACT: The purpose of this study was to identify genes that were differentially expressed in radiation-sensitive, naïve HL60 cells, and the derivatives created in our laboratory as indicated in the 'sample title'. We wanted to identify genes that were differentially expressed both before and after ionizing radiation (IR) exposure, from both a single dose of gamma rays and a single dose of alpha particles, at 4h following IR exposure. The data were used to identify genes that could be driving radioresistance in each respective cell line. The four cell lines, HL60, RA11, RG8, and RV+ were all analyzed at control (0Gy) radiation dose, 8Gy gamma rays, and ~2Gy alpha particles. Samples were collected from each cell line, for each dose, 4h following radiation exposure. The samples were collected from three independent experiments from three consecutive days in cell culture.

ORGANISM(S): Homo sapiens

SUBMITTER: Kurtis Haro

PROVIDER: E-GEOD-35372 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The growing number of particle treatment facilities worldwide and patients treated with particles instead of X-rays marks the upcoming rearrangement of modern radiotherapy. Especially for tumors being difficult to access and for tumors that are resistant to conventional X-ray treatment particle radiotherapy is a beneficial technology. At the Heidelberg Ion Beam Therapy Center (HIT) patients are treated with this technology since 2009 as it offers clear benefits. Contrary to X-rays, which show an exponential dose decrease (after reaching electron equilibrium) with increasing tissue depth, charged particles deposit most of their energy to a small region within the tissue with a sharp dose fall-off after the so-called Bragg peak. This precise dose localization enables further dose escalation within the tumor while sparing healthy tissue. Besides physical advantages, particle radiotherapy offers additional biological advantages. In radiobiology, the term relative-biological effectiveness (RBE) is defined as the ratio of X-rays dose to an alternative irradiation modality dose which produce the same biological effect (e.g. survival or number of DSBs). While protons have a relative biological effectiveness (RBE) comparable to X-rays, carbon ions are more effective in inducing DNA damage(1, 2) and are therefore especially useful for radioresistant tumors. This is due to the fact, that carbon ions induce clustered and direct DNA damage, which is considered to be less dependent on cell cycle stage, oxygen level, genetic background and hinders DNA repair mechanisms(3–6). Nevertheless, their exact mode of action and cellular mechanisms are largely unknown. We show the first comprehensive proteomic and phosphoproteomic study elucidating the cellular response to treatment with protons, carbon ions and X-rays. We found that 2h after treatment with these radiations negligible regulation occurred at protein expression level. But 181 phosphorylation sites were deregulated by ionizing radiation contributing mainly to DNA damage response functionalities. Interestingly we found 55 phosphorylation sites being differentially regulated between the radiations. Here, we observed protons and carbon ions producing equal cellular response whereas X-rays show altered regulation for certain phosphorylation sites. A subset of 28 phosphorylation sites being involved in the DNA damage response or differentially regulated between the ionizing radiations was selected for result confirmation.

Dataset Information

Genome-wide gene expression analysis of HL60 cells and its alpha particle-resistant derivative RA11, gamma resistant-derivative RG8, and vincristine resistant-derivative RV+ at baseline and following 8Gy gamma IR and ~2Gy alpha IR

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