中国医学科学院学报
中國醫學科學院學報
중국의학과학원학보
ACTA ACADEMIAE MEDICINAE SINICAE
2009年
6期
686-691
,共6页
凋亡%细胞色素c氧化酶%活性氧
凋亡%細胞色素c氧化酶%活性氧
조망%세포색소c양화매%활성양
apoptosis%cytochrome c oxidase%reactive oxygen species
目的 探讨亚硒酸钠诱导NB4细胞凋亡的过程中,细胞色素c氧化酶亚基IV(COX IV)表达的变化情况及其机制和意义.方法 用20μmol/L亚硒酸钠作用NB4细胞不同时间,Western blot和RT-PCR分别检测COX IV蛋白和mRNA表达的变化.活性氧清除剂预处理细胞后Western blot检测COX IV蛋白表达的变化以及caspase-3的激活.caspase-3抑制剂预处理细胞后Western blot检测COX IV蛋白表达的变化.瞬时转染干扰NB4细胞中COX IV表达后,流式细胞术检测细胞凋亡.结果 亚硒酸钠诱导NB4细胞COX IV蛋白表达显著下调,但mRNA表达水平无变化.活性氧清除剂能完全逆转亚硒酸钠引起的COX IV表达下调与caspase-3激活.caspase-3抑制剂能部分逆转亚硒酸钠引起的COX IV表达下调.干扰COX IV表达后,亚硒酸钠诱导的细胞凋亡显著增加.结论 在亚硒酸钠诱导NB4细胞凋亡过程中,COX IV在蛋白水平显著下调.活性氧介导COX IV下调与caspase-3激活,caspase-3在一定程度上参与COX IV表达下调,抑制COX IV能显著提高NB4细胞对亚硒酸钠诱导凋亡的敏感性.
目的 探討亞硒痠鈉誘導NB4細胞凋亡的過程中,細胞色素c氧化酶亞基IV(COX IV)錶達的變化情況及其機製和意義.方法 用20μmol/L亞硒痠鈉作用NB4細胞不同時間,Western blot和RT-PCR分彆檢測COX IV蛋白和mRNA錶達的變化.活性氧清除劑預處理細胞後Western blot檢測COX IV蛋白錶達的變化以及caspase-3的激活.caspase-3抑製劑預處理細胞後Western blot檢測COX IV蛋白錶達的變化.瞬時轉染榦擾NB4細胞中COX IV錶達後,流式細胞術檢測細胞凋亡.結果 亞硒痠鈉誘導NB4細胞COX IV蛋白錶達顯著下調,但mRNA錶達水平無變化.活性氧清除劑能完全逆轉亞硒痠鈉引起的COX IV錶達下調與caspase-3激活.caspase-3抑製劑能部分逆轉亞硒痠鈉引起的COX IV錶達下調.榦擾COX IV錶達後,亞硒痠鈉誘導的細胞凋亡顯著增加.結論 在亞硒痠鈉誘導NB4細胞凋亡過程中,COX IV在蛋白水平顯著下調.活性氧介導COX IV下調與caspase-3激活,caspase-3在一定程度上參與COX IV錶達下調,抑製COX IV能顯著提高NB4細胞對亞硒痠鈉誘導凋亡的敏感性.
목적 탐토아서산납유도NB4세포조망적과정중,세포색소c양화매아기IV(COX IV)표체적변화정황급기궤제화의의.방법 용20μmol/L아서산납작용NB4세포불동시간,Western blot화RT-PCR분별검측COX IV단백화mRNA표체적변화.활성양청제제예처리세포후Western blot검측COX IV단백표체적변화이급caspase-3적격활.caspase-3억제제예처리세포후Western blot검측COX IV단백표체적변화.순시전염간우NB4세포중COX IV표체후,류식세포술검측세포조망.결과 아서산납유도NB4세포COX IV단백표체현저하조,단mRNA표체수평무변화.활성양청제제능완전역전아서산납인기적COX IV표체하조여caspase-3격활.caspase-3억제제능부분역전아서산납인기적COX IV표체하조.간우COX IV표체후,아서산납유도적세포조망현저증가.결론 재아서산납유도NB4세포조망과정중,COX IV재단백수평현저하조.활성양개도COX IV하조여caspase-3격활,caspase-3재일정정도상삼여COX IV표체하조,억제COX IV능현저제고NB4세포대아서산납유도조망적민감성.
Objective To explore the mechanism and significance of cytochrome c oxidase subunit IV (COX IV) downregulation during apoptosis of NB4 cells induced by sodium selenite. MethodsNB4 cells were treated with 20 μmol/L sodium selenite at different time points. COX IV protein and mRNA were detected by Western blot and RT-PCR, respectively. NB4 cells were pretreated with reactive oxygen species (ROS) scavenger before selenite exposure, and then COX IV protein expression and caspase-3 activation were detected by Western blot. NB4 cells were pretreated with caspase-3 inhibitor before selenite exposure, and then COX IV protein expression was detected by Western blot. NB4 cells were transiently transfected with vectors to interfere with the expression of COX IV, and then the apoptosis induced by selenite was analyzed by flow cytometry. ResultsSodium selenite induced evident downregulation of COX IV protein in NB4 cells, while its mRNA level was almost unchanged. ROS scavenger completely reversed selenite-induced COX IV downregulation and caspase-3 activation. Caspase-3 inhibitor partially reversed selenite-induced COX IV downregulation. Interference with COX IV expression dramatically enhanced selenite-induced apoptosis of NB4 cells. ConclusionsCOX IV is remarkaly downregulated during selenite-induced apoptosis of NB4 cells. ROS mediates COX IV downregulation and caspase-3 activation, while caspase-3 is partially involved in COX IV downregulation. COX IV interference markedly increases the sensitivity of NB4 cells to selenite-induced apoptosis.
