The Effect of Radiotherapy on Cell Survival and Inflammatory Cytokine and Chemokine Secretion in a Co-Culture Model of Head and Neck Squamous Cell Carcinoma and Normal Cells.

Radiotherapy (RT) is one of the main treatments for head and neck squamous cell carcinomas (HNSCCs). Unfortunately, radioresistance is observed in many cases of HNSCCs. The effectiveness of RT depends on both the direct effect inducing cell death and the indirect effect of changing the tumor microenvironment (TME). Knowledge of interactions between TME components after RT may help to design a new combined treatment with RT. In the study, we investigated the effect of RT on cell survival and cell secretion in a co-culture model of HNSCCs in vitro. We examined changes in cell proliferation, colony formation, cell cycle phases, type of cell death, cell migration and secretion after irradiation. The obtained results suggest that the presence of fibroblasts and endothelial cells in co-culture with HNSCCs inhibits the function of cell cycle checkpoints G1/S and G2/M and allows cells to enter the next phase of the cell cycle. We showed an anti-apoptotic effect in co-culture of HNSCCs with fibroblasts or endothelial cells in relation to the execution phase of apoptosis, although we initially observed increased activation of the early phase of apoptosis in the co-cultures after irradiation. We hypothesize that the anti-apoptotic effect depends on increased secretion of IL-6 and MCP-1.

NMR-Based Metabolomics in Investigation of the Radiation Induced Changes in Blood Serum of Head and Neck Cancer Patients and Its Correlation with the Tissue Volumes Exposed to the Particulate Doses.

In the present study, we analyze the nuclear magnetic resonance (NMR) blood serum metabolic profiles of 106 head and neck squamous cell carcinoma (HNSCC) patients during radio (RT) and concurrent radio-chemotherapy (CHRT). Four different fractionation schemes were compared. The blood samples were collected weekly, from the day before the treatment until the last week of CHRT/RT. The NMR spectra were acquired on A Bruker 400 MHz spectrometer at 310 K and analyzed using multivariate methods. Seven metabolites were found significantly to be altered solely by radiotherapy: N-acetyl-glycoprotein (NAG), N-acetylcysteine, glycerol, glycolate and the lipids at 0.9, 1.3 and 3.2 ppm. The NMR results were correlated with the tissue volumes receiving a particular dose of radiation. The influence of the irradiated volume on the metabolic profile is weak and mainly limited to sparse correlations with the inflammatory markers, creatinine and the lymphocyte count in RT and the branched-chain amino-acids in CHRT. This is probably due to the optimal planning and delivery of radiotherapy improving sparing of the surrounding normal tissues and minimizing the differences between the patients (caused by the tumor location and size).

Dose-dependence of radiotherapy-induced changes in serum levels of choline-containing phospholipids; the importance of lower doses delivered to large volumes of normal tissues.

BACKGROUND: Conformal radiotherapy is a primary treatment in head and neck cancer, which putative adverse effects depend on relatively low doses of radiation delivered to increased volumes of normal tissues. Systemic effects of such treatment include radiation-induced changes in serum lipid profile, yet dose- and volume-dependence of these changes remain to be established. METHODS: Here we analyzed levels of choline-containing phospholipids in serum samples collected consecutively during the radiotherapy used as the only treatment modality. The liquid chromatography-mass spectrometry (LC-MS) approach applied in the study enabled the detection and quantitation of 151 phospholipids, including (lyso)phosphatidylcholines and sphingomyelins. RESULTS: No statistically significant differences were found in the pretreatment samples from patients with different locations and stages of cancer. To compensate for potential differences between schemes of radiotherapy, the biologically effective doses were calculated and used in the search of correlations with specific lipid levels. We found that the levels of several phospholipids depended on the maximum dose delivered to the gross tumor volume and total radiation energy absorbed by the patient's body. Increased doses correlated with increased levels of sphingomyelins and reduced levels of phosphatidylcholines. Furthermore, we observed several phospholipids whose serum levels correlated with the degree of acute radiation toxicity. CONCLUSION: Noteworthy, serum phospholipid levels were associated mainly with volumes of normal tissues irradiated with relatively low doses (i.e., total accumulated dose 20 Gy), which indicated the importance of such effects on the systemic response of the patient's organism to intensity-modulated radiotherapy (IMRT).

[Comparison of two image guided radiation therapy (IGRT) methods used for prostate cancer patients--CBCT and 2D-2D kV]. / Porównanie dwóch metod radioterapii sterowanej obrazem (IGRT) chorych na raka stercza--CBCT i 2D-2D kV.

The IGRT notion (image guided radiation therapy) comprises all techniques enabling checking and correction of patients position directly before or during an irradiation session. In last years they became a standard in radiotherapy due to decreasing of geographical misses. The aim of our study was a comparison of two IGRT techniques--CBCT and 2D-2D kV performed for prostate cancer patients and a comparison of two immobilization systems used for them. The performed analysis comprises 3582 2D-2D kV and 2110 CBCT IGRT measurements made for 85 prostate cancer patients. Patients were irradiated using thermoplastic masks and two kinds of immobilizing plates. One subgroup of patients was treated using leg supports and second one without them. Mean, maximum, minimum and standard deviation of absolute values of shifts (cm) measured using 2D-2D kV were 0.27, 0.0, 2.8 and 0.33 respectively. For CBCT these same values were 0.31, 0.0, 4.1 and 0.33. Mean, maximum, minimum and standard deviation of real shifts values (cm) measured using 2D-2D kV were -0.01, -2.5, 2.8 and 0.43 respectively. For CBCT these same values were 0.01, -4.1, 2.3 and 0.45. Comparison of shift absolute values distributions for whole analyzed group showed statistically significant difference (p=0.001) between 2D-2D kV and CBCT with higher mean for CBCT (0.31 vs 0.27) and equal standard deviations. Statistically significant difference (p=0.000...) between distributions of measurements in z axis was found (for 2D-2D kV mean 0.16, SD=0.21, for CBCT mean 0.25, SD=0.25). Comparison of absolute shifts values distributions revealed significant difference for CBCT, for two immobilization plates--in x (p=0.001) and y axis (p=0.007). For the small plate in x axis mean was 0.22 (SD=0.2), and for the large one (with an integrated head support) 0.17 (SD=0.2). Comparison of absolute shifts values for sub- groups created dependently on the leg support use, showed significant differences in x axis for 2D-2D kV (p=0.000...); mean in the subgroup irradiated without leg supports was 0.14 (SD=0.14) and for the subgroup without them was 0.19 (SD=0.15) and in x axis for CBCT (p=0.03); mean in the subgroup irradiated without leg supports was 0.18 (SD=0.18) and for the subgroup without them was 0.23 (SD=0.22). Significant difference was also revealed for the whole group of absolute shift values for 2D-2D kV (p=0.01). For the subgroup irradiated without leg supports mean was 0.27 (SD=0.34), and for the subgroup treated with them was 0.27 (SD=0.31). Obtained results permit us to conclude that in the case of prostate cancer patients IGRT based on the bone anatomy visualization, the method of choice should be 2D-2D kV, because it allows for more precise and shorter patient position evaluation and, that during IGRT of prostate cancer patients using a simple thermoplastic immobilization system is sufficient; use of more sophisticated systems with additional supports did not improve the patient immobilization.