线粒体DNA缺失对人淋巴瘤Namalwa细胞生长和侵袭能力的影响

Effect of mitochondrial DNA deletion on growth and invasiveness of human lymphoma Namalwa cells

目的探讨线粒体DNA(mtDNA)缺失对人淋巴瘤Namalwa细胞生长和迁移能力的影响。方法采用溴化乙锭处理Namalwa细胞获得mtDNA缺失的ρ0-Namalwa细胞;营养缺陷法、PCR扩增线粒体DNA片段、Western bloting检测COXII蛋白表达等方法鉴定ρ0-Namalwa细胞;MTT方法及细胞周期测定细胞增殖能力;Transwell实验检测细胞迁移及侵袭能力;流式细胞术检测活性氧(ROS)及细胞内钙离子水平。结果ρ0-Namalwa细胞在选择性培养基中正常生长,而在非选择性培养基中逐渐死亡;ρ0-Namalwa细胞未扩增出mtDNA片段以及未表达COXII蛋白;与Namalwa细胞相比,ρ0-Namalwa的细胞增殖率、迁移及侵袭能力、ROS水平及细胞内Ca2+浓度明显降低,细胞阻滞于G0/G1期。结论ρ0-Namalwa细胞与亲本Namalwa细胞相比,生长及迁移能力减弱,可能与细胞内ROS产生减少及细胞内Ca2+浓度降低有关。

Objective To explore the effect of mitochondrial DNA （mtDNA） deletion on the growth and invasiveness of human lymphoma Namalwa cells. Methods p°-Namalwa cells with mtDNA deletion were generated by treating Namalwa cells with ethidium bromide and confirmed by selective p° test medium analysis, PCR and Western blotting. The growth of p°-Namalwa cells was evaluated by MTT assay and cell cycle analysis, and the cell migration and invasiveness were assessed with Transwell assay. Reactive oxygen species （ROS） production and cytosolic Ca2＋ were detected by flow cytometry. Results p°-Namalwa cells could grow and divide normally in selective medium supplemented with uridine and pyruvate but not in nonselective medium. PCR did not yield the products of mtDNA, nor was COXII expression detected in p°-Namalwa cells, p°-Namalwa cells showed an obvious attenuation of cell proliferation and migration abilities with significantly lowered ROS production and cytosolic Ca^2＋. Conclusion The suppressed growth and migration of p°-Namalwa cells may be the result of decreased ROS production and cytosolic Ca^2＋