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目的观察CD20单抗美罗华(R)对X射线所致人Burkitt淋巴瘤细胞系Raji细胞损伤的影响及与细胞内活性氧(ROS)的关系。方法Raji细胞与低浓度H_2O_2或美罗华(终浓度20mg/L)孵育24h后,给予X射线照射;Raji细胞经美罗华联合不同浓度H_2O_2处理,常规培养48h,采用SRB法检测细胞存活,AO/EB双染观察细胞凋亡变化,激光共聚焦显微镜动态检测细胞内活性氧水平。结果联合美罗华后,Raji细胞对X射线诱导的细胞死亡分数明显提高,美罗华+照射组和照射组的ID50分别为2.80 Gy、5.08 Gy;20μM或40μM的低剂量H_2O_2也可以提高Raji细胞对辐射的敏感性,并呈现浓度依赖性。与单照射组相比较,美罗华+照射组在处理后24h,出现明显细胞凋亡,可被抗氧化剂NAC部分抑制。美罗华联合照射组ROS产生明显高于单照射组,同时,美罗华可增加Raji细胞对不同浓度H_2O_2所致氧胁迫的敏感性,表现为细胞生长抑制和凋亡诱导增加。结论美罗华可以增加耐辐射的淋巴瘤细胞系Raji细胞对X射线的敏感性,ROS作为细胞内外重要的信号分子,可能发挥重要作用。
Objective To investigate the effect of CD20 monoclonal antibody Rituximab on the injury of Raji cells induced by X-ray in human Burkitt’s lymphoma cell line and its relationship with intracellular reactive oxygen species (ROS). Methods Raji cells were incubated with low concentration of H 2 O 2 or rituximab (final concentration of 20 mg / L) for 24 h. The Raji cells were treated with rituximab and different concentrations of H 2 O 2 for 48 h. The survival rate of Raji cells was measured by SRB. The changes of apoptosis were observed and the levels of reactive oxygen species (ROS) in cells were detected by laser confocal microscopy. Results After combined with Rituximab, Raji cells significantly increased the X-ray-induced cell death scores. The ID50 of rodenticide + irradiation group and irradiation group were 2.80 Gy and 5.08 Gy, respectively. Low doses of 20 μM or 40 μM H_2O_2 could also increase Raji cell radiation Sensitivity, and concentration-dependent. Compared with the single irradiation group, the rituximab + irradiation group showed obvious apoptosis 24 h after treatment, and could be partially inhibited by the antioxidant NAC. ROS production in the rituximab group was significantly higher than that in the single irradiation group. In addition, rituximab increased the sensitivity of Raji cells to different concentrations of H 2 O 2 -induced oxygen stress, which showed that the cell growth was inhibited and the induction of apoptosis increased. Conclusion Rituximab can increase the X-ray sensitivity of Raji cells, a radiation-tolerant lymphoma cell line. ROS may play an important role as an important signal molecule both inside and outside cells.