Rituximab enhances radiation-triggered apoptosis in non-Hodgkin's lymphoma cells via caspase-dependent and - independent mechanisms.

Rituximab (RTX), a chimeric human anti-CD20 monoclonal antibody, is currently employed in the treatment of malignant non-Hodgkin's lymphoma (NHL) either alone or in combination with other cytotoxic approaches. The present study examines the effects of ionizing radiation in combination with RTX on proliferation and apoptosis development in B-lymphoma RL and Raji cells. RTX was used at a concentration of 10 microg/mL 24 hours prior to irradiation at a single dose of 9 Gy. CD20 expression, cell viability, apoptosis, mitochondrial membrane potential and apoptosis-related proteins were evaluated in the treated B cells. The constitutive level of CD20 expression in RL and Raji lymphoma cells did not play an essential role in RTX-induced cell growth delay. Both lymphoma cells showed similar inhibition of cell proliferation without apoptosis development in response to RTX treatment. Exposure to ionizing radiation induced cell growth delay and apoptosis in RL cells, whereas Raji cells showed moderate radio-resistance and activation of cell growth at 24 hours after irradiation, which was accompanied by increased radiation-triggered CD20 expression. The simultaneous exposure of lymphoma cells to ionizing radiation and RTX abrogated radioresistance of Raji cells and significantly enhanced cell growth delay and apoptosis in RL cells. X-linked inhibitor of apoptosis protein (XIAP) and the inducible form of heat shock protein 70 (Hsp70) were positively modulated by RTX in combination with ionizing radiation in order to induce apoptosis. Furthermore, it was demonstrated that mitochondrial membrane potential dissipation is not an essential component to induce apoptosis-inducing factor (AIF) maturation and apoptosis. Our results show that RTX-triggered enhancement of radiation-induced apoptosis and cell growth delay is achieved by modulation of proteins involved in programmed cell death.

Pretreatment with rituximab enhances radiosensitivity of non-Hodgkin's lymphoma cells.

The present study examines the effects of ionizing radiation in combination with rituximab (RTX), a chimeric human anti-CD20 monoclonal antibody, on proliferation, cell cycle distribution and apoptosis in B-lymphoma RL and Raji cells. Exposure to ionizing radiation (9 Gy) induced cell growth delay and apoptosis in RL cells, whereas Raji cells showed moderate radio-resistance. The simultaneous exposure of lymphoma cells to ionizing radiation and RTX (10 microg/mL) markedly enhanced apoptosis and cell growth delay in RL and Raji cells. Cooperative antiproliferative and apoptotic effects of RTX and radiation were achieved through the inhibition of c-myc and bcl-XL expression. Furthermore, RTX-modulated expression of cell cycle regulating proteins, such as p53, p21/WAF1, p27/KIP1, contributed to the development of radiation-induced cell killing and growth arrest. Each NHL cell line that underwent apoptosis induced by combination treatment revealed enhanced caspase-3 and poly (ADP-ribose) polymerase (PARP) cleavage as compared to only irradiated cells. These findings show that rituximab synergistically enhances radiation-induced apoptosis and cell growth delay through the expression of proteins involved in the programmed cell death and cell cycle regulation pathways.

Competing irradiation techniques for para-aortic lymph nodes: dose distribution and NTCP for the kidney.

PURPOSE: Partial coirradiation of both kidneys is an unavoidable consequence of adequate dose delivery in radiation therapy of para-aortic lymph nodes (PLN). Depending on total dose anteroposterior/posteroanterior (AP/PA), opposed-fields or multifield techniques are used. To optimize the treatment of potentially tumor-affected PLN with minimal kidney involvement, we calculated normal tissue complication probabilities (NTCPs) of coirradiated kidneys for four common irradiation techniques used in the PLN area. METHODS AND MATERIALS: Planning target volume (PTV) delineation was performed in computed tomography scans of 21 patients with a lateral safety margin of 3 cm from the aorta and 2 cm aside the vena cava. Ventral and dorsal margins of the PTV were delineated 2 cm from the vessels. As previously shown (Nevinny-Stickel M, et al. Int J Radiat Oncol Biol Phys 2000;48:147-151), PTVs optimized by these altered delineations permit inclusion of at least 97% of potentially involved PLN in contrast to standard delineations based on bony structures that are more likely to miss affected lymph nodes. The present study compared NTCPs for individual PTV-based treatment planning with NTCPs for standard planning based on bony structures. For each patient, four hypothetical treatment plans were created: (A) standard AP/PA opposed fields technique with lateral field margins along the tips of the transverse processes of the vertebral bodies; (B) individually planned AP/PA opposed fields with lateral field margins according to the optimized PTV; (C) standard four-field box technique with lateral width as described for (A), with dorsal borders at the center of the vertebral bodies and ventral margins 3 cm in front of the vertebrae; and (D) individually planned four-field box with lateral field margins according to the optimized PTV. Calculation of irradiation-induced complication probability values for nonuniform kidney irradiation was performed for model doses 19.8 Gy, 30.6 Gy, and 50.4 Gy according to the Lyman-Wolbarst model. RESULTS: No dose showed a statistically significant difference (p < 0.00833, corrected for six multiple interrelated comparisons) in the median of total organ kidney NTCPs between techniques A, C, and D, with technique D intermediately ranging between technique A and C (e.g., for 50.4 Gy: A: median, 0.39; range, 0.01-0.83; C: median, 0.27 range; 0.05-0.68; D: 0.36; range, 0.03-0.72). In comparison to techniques A, C, and D, the individually planned AP/PA opposed-fields technique (B) was accompanied by significantly higher and intolerable overall kidney NTCP rates (e.g., for 50.4 Gy: median, 0.68; range, 0.01-0.99). CONCLUSION: Conformal four-field planning with individually optimized PTVs (D) resulted in only moderate tissue complication probabilities in both kidneys with the advantage of providing significantly greater inclusion of potentially involved PLNs in comparison to accepted standard procedures (A and C).