辐射诱导小鼠淋巴瘤细胞凋亡涉及NADPH氧化酶的活性

γ-Irradiation Induced Mouse Lymphoma Cell Apoptosis Involved the NADPH Oxidase Activity

用60Coγ射线对小鼠淋巴瘤细胞3SB(p53+/+)和1B1C4(p53-/-)进行照射处理。观察了辐射后细胞形态学变化,结果发现,3SB对γ射线诱导的凋亡非常敏感,而1B1C4细胞对辐射具有抗性。DNA片断化试验和藻红B染色试验的结果也证实这一点。进一步分析了凋亡信号途径上的caspase-3和p53。在3SB细胞中,检测到激活的caspase-3并发现p53的表达随辐射剂量的增加而增高。比较了两种细胞中caspase-3mRNA的表达,但未见差异。对两种细胞中的NADPH氧化酶活性的测定结果表明,凋亡的3SB细胞中的NADPH氧化酶活性增高。因此,在电离辐射诱导细胞凋亡的过程中,NADPH氧化酶参与了上游信号的转导。

Mouse Lymphoma cell 3SB （p53^＋/＋） and 1B1C4 （p53^-/-） were irradiated with ^60Co γ-ray. Their apoptotic morphology was observed with microscope. Apoptotic assay results demonstrated that those two cell lines were quite differently to γ-irradiation. 3SB was very sensitive, while 1B1C4 was anti-sensitive to γ-irradiation. Both DNA ladder experiment and erythrosine B staining results confirmed this conclusion. The signal transduction molecular caspase-3 and p53 were also analyzed. The activated caspase-3 and p53 were found up-regulated by γ-irradiation with dose pattern in 3SB cell. The caspase-3 mRNA differential expressions were also amplified between those two cell lines, but no difference was found. Then the activity of NADPH oxidase was checked in these two cells, the activity of NADPH oxidase was found only in apoptotic 3SB cell. In our research we found that NADPH-oxidase is another upstream signal molecular which involved in irradiation induced apoptosis.