南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (02): 188-.

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线粒体DNA缺失对人淋巴瘤Namalwa 细胞生长和侵袭能力的影响

马佳，张庆，陈昌杰，杨清玲，陈素莲   

  • 出版日期:2013-02-20 发布日期:2013-02-20

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

  • Online:2013-02-20 Published:2013-02-20

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摘要： ：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。

Abstract: Objective To explore the effect of mitochondrial DNA (mtDNA) deletion on the growth and invasiveness of human
lymphoma Namalwa cells. Methods ρ0-Namalwa cells with mtDNA deletion were generated by treating Namalwa cells with
ethidium bromide and confirmed by selective ρ0 test medium analysis, PCR and Western blotting. The growth of ρ0-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 ρ0-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 ρ0-Namalwa cells. ρ0-Namalwa cells
showed an obvious attenuation of cell proliferation and migration abilities with significantly lowered ROS production and
cytosolic Ca2 + . Conclusion The suppressed growth and migration of ρ0-Namalwa cells may be the result of decreased ROS
production and cytosolic Ca2+.