中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2014年
3期
197-202
,共6页
柯跃斌%吴双%黄娟%袁建辉%邓平建%程锦泉
柯躍斌%吳雙%黃娟%袁建輝%鄧平建%程錦泉
가약빈%오쌍%황연%원건휘%산평건%정금천
RNA干扰%基因缺陷%DNA氧化%修复基因%生物标志%替补作用
RNA榦擾%基因缺陷%DNA氧化%脩複基因%生物標誌%替補作用
RNA간우%기인결함%DNA양화%수복기인%생물표지%체보작용
RNA interference%Genetic defect%DNA oxidation%Repair gene%Biomarker%Substitution effect
目的 通过建立基因缺陷细胞模型,探讨修复基因hOGG1与hMTH1在DNA氧化性损伤中的作用与关系.方法 利用慢病毒感染人胚肺成纤维细胞(HFL),分别建立hOGG1基因和hMTH1基因缺陷细胞模型.将HFL细胞在100 μmol/L的H2O2中孵育12h,分别利用高压液相色谱串联电化学检测技术及RT-PCR技术分析8-羟基-脱氧鸟嘌呤(7,8-dihydro-8-oxoguanine,8-oxo-dG)水平及hOGG1和hMTH1基因表达水平.结果 用高滴度慢病毒感染靶细胞后得到hOGG1基因和hMTH1基因缺陷细胞模型,hOGG1 mRNA表达水平(0.09±0.02)比正常HFL细胞(1.00±0.04)下降了91％,hMTH1 mRNA表达水平(0.41±0.04)比正常HFL细胞(1.02±0.06)下降了60％；用100 μmol/L的H2O2诱导12 h,hOGG1基因缺陷细胞的hMTH1基因表达水平(1.26±0.18)相比对照组(1.01±0.07)提高了25％,hMTH1基因缺陷细胞的hOGG1基因表达水平(1.54±0.25)提高了52％；hOGG1基因缺陷细胞的8-oxo-dG水平(2.48±0.54)是对照组(0.80±0.16)的3.1倍,差异有统计学意义(P＜0.01),hMTH1基因缺陷细胞(1.84±0.46)的8-oxo-dG水平是对照组(0.80±0.16)的2.3倍,差异均有统计学意义(P＜0.01).结论 利用基因缺陷细胞模型,发现在氧化诱导作用下,修复基因hOGG1与hMTH1在DNA损伤修复活动中可能分别表现出一定的协同性替补作用,hOGG1的替补作用强于hMTH1.
目的 通過建立基因缺陷細胞模型,探討脩複基因hOGG1與hMTH1在DNA氧化性損傷中的作用與關繫.方法 利用慢病毒感染人胚肺成纖維細胞(HFL),分彆建立hOGG1基因和hMTH1基因缺陷細胞模型.將HFL細胞在100 μmol/L的H2O2中孵育12h,分彆利用高壓液相色譜串聯電化學檢測技術及RT-PCR技術分析8-羥基-脫氧鳥嘌呤(7,8-dihydro-8-oxoguanine,8-oxo-dG)水平及hOGG1和hMTH1基因錶達水平.結果 用高滴度慢病毒感染靶細胞後得到hOGG1基因和hMTH1基因缺陷細胞模型,hOGG1 mRNA錶達水平(0.09±0.02)比正常HFL細胞(1.00±0.04)下降瞭91％,hMTH1 mRNA錶達水平(0.41±0.04)比正常HFL細胞(1.02±0.06)下降瞭60％；用100 μmol/L的H2O2誘導12 h,hOGG1基因缺陷細胞的hMTH1基因錶達水平(1.26±0.18)相比對照組(1.01±0.07)提高瞭25％,hMTH1基因缺陷細胞的hOGG1基因錶達水平(1.54±0.25)提高瞭52％；hOGG1基因缺陷細胞的8-oxo-dG水平(2.48±0.54)是對照組(0.80±0.16)的3.1倍,差異有統計學意義(P＜0.01),hMTH1基因缺陷細胞(1.84±0.46)的8-oxo-dG水平是對照組(0.80±0.16)的2.3倍,差異均有統計學意義(P＜0.01).結論 利用基因缺陷細胞模型,髮現在氧化誘導作用下,脩複基因hOGG1與hMTH1在DNA損傷脩複活動中可能分彆錶現齣一定的協同性替補作用,hOGG1的替補作用彊于hMTH1.
목적 통과건립기인결함세포모형,탐토수복기인hOGG1여hMTH1재DNA양화성손상중적작용여관계.방법 이용만병독감염인배폐성섬유세포(HFL),분별건립hOGG1기인화hMTH1기인결함세포모형.장HFL세포재100 μmol/L적H2O2중부육12h,분별이용고압액상색보천련전화학검측기술급RT-PCR기술분석8-간기-탈양조표령(7,8-dihydro-8-oxoguanine,8-oxo-dG)수평급hOGG1화hMTH1기인표체수평.결과 용고적도만병독감염파세포후득도hOGG1기인화hMTH1기인결함세포모형,hOGG1 mRNA표체수평(0.09±0.02)비정상HFL세포(1.00±0.04)하강료91％,hMTH1 mRNA표체수평(0.41±0.04)비정상HFL세포(1.02±0.06)하강료60％；용100 μmol/L적H2O2유도12 h,hOGG1기인결함세포적hMTH1기인표체수평(1.26±0.18)상비대조조(1.01±0.07)제고료25％,hMTH1기인결함세포적hOGG1기인표체수평(1.54±0.25)제고료52％；hOGG1기인결함세포적8-oxo-dG수평(2.48±0.54)시대조조(0.80±0.16)적3.1배,차이유통계학의의(P＜0.01),hMTH1기인결함세포(1.84±0.46)적8-oxo-dG수평시대조조(0.80±0.16)적2.3배,차이균유통계학의의(P＜0.01).결론 이용기인결함세포모형,발현재양화유도작용하,수복기인hOGG1여hMTH1재DNA손상수복활동중가능분별표현출일정적협동성체보작용,hOGG1적체보작용강우hMTH1.
Objective To investigate the potential substitution effect of hOGG1 and hMTH1 on oxidative DNA damage,based on gene-deficient cell strains models.Methods hOGG1 and hMTH1 gene deficient cell strains models were established by Human embryonic lung fibroblasts(HFL) cells.After HFL cells being exposed to 100 μmol/L H2O2 for 12 h,HPLC-EC detecting technique and RT-PCR method were adopted to analyze the genetic expression level of 8-oxo-dG (7,8-dihydro-8-oxoguanine).Results The gene-deficient cell strains models of hOGG1 and hMTH1 were obtained by infecting target cells with high titer of lentivirus.The mRNA expression level of hOGG1 was 0.09 ±0.02,91％ lower than it in normal HFL cells,which was 1.00 ± 0.04.As the same,the mRNA expression level of hMTH1 (0.41 ± 0.04) also decreased by 60％ compared with it in normal HFL cells (1.02 ± 0.06).After induced by 100 μmol/L H2O2 for 12 h,the genetic expression level of hMTH1 in hOGG1 gene-deficient cells (1.26 ± 0.18) increased 25％ compared with it in control group (1.01 ± 0.07).Meanwhile,the genetic expression level of hOGG1 in hMTH1 gene-deficient cells (1.54 ± 0.25) also increased by 52％.The DNA 8-oxo-dG levels in hOGG1 gene-deficient cells (2.48 ± 0.54) was 3.1 times compared with it in the control group (0.80 ± 0.16),the difference showed statistical significance (P ＜0.01).Whereas the 8-oxo-dG levels in hMTH1 gene-deficient cells (1.84 ± 0.46) was 2.3 times of it in the control group,the difference also showed statistical significance (P ＜ 0.01).Conclusion Based on gene-deficient HFL cells models,a synergetic substitution effect on DNA damage and repair activity by both hOGG1 and hMTH1 were firstly discovered when induced by oxidation.The substitution effect of hOGG1 were stronger than that of hMTH1.
