肝癌患者单个核细胞线粒体DNA拷贝数与抗氧化能力的变化  

Changes of the mitochondrial DNA copy number and the antioxidant system in the PBMC of hepatocellular carcinoma

作　　者：高园[1] 聂鸿靖[1] 杨栋[1] 丁诚实[1,2] 金敏[1] 谌志强[1] 邱志刚[1] 郭旋[1] 陈照立[1] 李君文[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050 [2]枣庄学院,山东枣庄277160

出　　处：《中国应用生理学杂志》2016年第1期1-5,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(81471812)

摘　　要：目的:探讨肝癌患者单个核细胞(PBMC)线粒体DNA(mt DNA)拷贝数的变化及其与机体抗氧化能力的关系。方法:用Ficoll Hypaque离心法分离外周血单个核细胞(PBMC),采用实时荧光定量PCR反应,检测线粒体NADH脱氢酶亚基1(ND1)基因的拷贝数,并以核基因组的β-actin作为内参基因,比较25例肝癌患者与30例健康人外周血单个核细胞中mt DNA拷贝数的差异。流式细胞术检测单个核细胞活性氧(ROS)含量的变化。生化检测法检测血浆中总抗氧化能力(T-AOC)的变化。结果:肝癌组外周血单个核细胞ND1基因拷贝数是健康对照组的73%,表明肝癌患者外周血单个核细胞mt DNA拷贝数明显下降。肝癌组单个核细胞活性氧含量的平均荧光强度为(417.82±110.62),健康对照组为(301.82±75.54),肝癌组单个核细胞活性氧含量显著高于健康对照组(P<0.01)。肝癌组血浆总抗氧化能力(单位/毫升血浆)吸光度为(1.30±0.85),健康对照组为(3.20±1.62),肝癌组血浆总抗氧化能力显著低于健康对照组(P<0.01)。结论:肝癌患者的外周血单个核细胞线粒体DNA拷贝数减少可能与机体抗氧化水平下降有关。Objective： To investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nuclear cell（PBMC） and the disordered of antioxidant capacity of hepatocelltdar carcinoma（HCC） patients. Methods： The Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The NDI gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species （ROS） in PBMC was determined by flow cytometry. The change of total antioxidant capacity （TAOC） of plasma was detected by the biochemistry examination. Results： The copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group, which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was （417. 82 ± 110.62） and （301.82 ± 75.54） in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group（ P 〈 0.01 ）. Plasma T-AOC in HCC was （ 1.30 ± 0.85 ）, and （3.20 ± 1.62） in control. The T-AOC of plasma of HCC was significantly lower than in control group（ P 〈 0.01 ）. Conclusion： There was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may beassociated with the development of hepatocellular carcinoma.

关 键 词：肝癌 线粒体DNA 活性氧 总抗氧化能力 单个核细胞 

分 类 号：R735.7[医药卫生—肿瘤]