中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2008年
7期
484-489
,共6页
藤黄酸%白血病%凋亡%Nup88
籐黃痠%白血病%凋亡%Nup88
등황산%백혈병%조망%Nup88
Gambogic acid%Leukemia%Apoptosis%Nup88
目的 观察藤黄酸对白血病细胞株HL-60增殖和凋亡的影响及其对核孔蛋白Nup88的调控作用,探讨二者间的相互关系.方法 采用二苯基溴化四氮唑蓝(MTT)比色法检测细胞增殖活性;Annexin-V FITC/PI双标法和Hoechst33258染色法分析细胞凋亡的改变;流式细胞术分析细胞周期和细胞内核孔蛋白Nup88的改变;逆转录聚合酶链反应(RT-PCR)检测藤黄酸对白血病细胞内Nup88基因表达的影响;共聚焦显微镜下观察Nup88蛋白的分布情况.结果 藤黄酸能明显抑制HL-60细胞的增殖,其抑制作用呈时间、剂量依赖性,其12 h的IC50Molecule targetecl therapy; Response evaluation criteria为1.797 μmol/L.藤黄酸具有较强的诱导白血病细胞凋亡的效应,0.4 μmol/L藤黄酸即能诱导15.1%的HL-60细胞发生凋亡.当藤黄酸浓度达到1.6 μmol/L时,总凋亡率为79.0%,且该效应并不依赖于细胞周期阻滞作用.核孔蛋白Nup88弥漫分布于白血病细胞的核浆之间,以细胞浆和核膜为主,经藤黄酸干预后,Nup88蛋白和mRNA的表达水平明显下降,主要集中于核膜的胞浆面,偶见胞浆中表达.结论 藤黄酸能明显抑制HL-60白血病细胞的增殖,并诱导其凋亡;藤黄酸诱导的核孔蛋白Nup88的重新分布和表达量的下调可能参与了其诱导凋亡作用.
目的 觀察籐黃痠對白血病細胞株HL-60增殖和凋亡的影響及其對覈孔蛋白Nup88的調控作用,探討二者間的相互關繫.方法 採用二苯基溴化四氮唑藍(MTT)比色法檢測細胞增殖活性;Annexin-V FITC/PI雙標法和Hoechst33258染色法分析細胞凋亡的改變;流式細胞術分析細胞週期和細胞內覈孔蛋白Nup88的改變;逆轉錄聚閤酶鏈反應(RT-PCR)檢測籐黃痠對白血病細胞內Nup88基因錶達的影響;共聚焦顯微鏡下觀察Nup88蛋白的分佈情況.結果 籐黃痠能明顯抑製HL-60細胞的增殖,其抑製作用呈時間、劑量依賴性,其12 h的IC50Molecule targetecl therapy; Response evaluation criteria為1.797 μmol/L.籐黃痠具有較彊的誘導白血病細胞凋亡的效應,0.4 μmol/L籐黃痠即能誘導15.1%的HL-60細胞髮生凋亡.噹籐黃痠濃度達到1.6 μmol/L時,總凋亡率為79.0%,且該效應併不依賴于細胞週期阻滯作用.覈孔蛋白Nup88瀰漫分佈于白血病細胞的覈漿之間,以細胞漿和覈膜為主,經籐黃痠榦預後,Nup88蛋白和mRNA的錶達水平明顯下降,主要集中于覈膜的胞漿麵,偶見胞漿中錶達.結論 籐黃痠能明顯抑製HL-60白血病細胞的增殖,併誘導其凋亡;籐黃痠誘導的覈孔蛋白Nup88的重新分佈和錶達量的下調可能參與瞭其誘導凋亡作用.
목적 관찰등황산대백혈병세포주HL-60증식화조망적영향급기대핵공단백Nup88적조공작용,탐토이자간적상호관계.방법 채용이분기추화사담서람(MTT)비색법검측세포증식활성;Annexin-V FITC/PI쌍표법화Hoechst33258염색법분석세포조망적개변;류식세포술분석세포주기화세포내핵공단백Nup88적개변;역전록취합매련반응(RT-PCR)검측등황산대백혈병세포내Nup88기인표체적영향;공취초현미경하관찰Nup88단백적분포정황.결과 등황산능명현억제HL-60세포적증식,기억제작용정시간、제량의뢰성,기12 h적IC50Molecule targetecl therapy; Response evaluation criteria위1.797 μmol/L.등황산구유교강적유도백혈병세포조망적효응,0.4 μmol/L등황산즉능유도15.1%적HL-60세포발생조망.당등황산농도체도1.6 μmol/L시,총조망솔위79.0%,차해효응병불의뢰우세포주기조체작용.핵공단백Nup88미만분포우백혈병세포적핵장지간,이세포장화핵막위주,경등황산간예후,Nup88단백화mRNA적표체수평명현하강,주요집중우핵막적포장면,우견포장중표체.결론 등황산능명현억제HL-60백혈병세포적증식,병유도기조망;등황산유도적핵공단백Nup88적중신분포화표체량적하조가능삼여료기유도조망작용.
Objective To investigate the effect of gambogic acid (GA) on cell proliferation and induction of apoptosis in HL-60 cells in vitro, as well as the regulation of nucleoporin Nup88 to explore the relationship between them. Methods The effect of GA on the growth of HL-60 cells was determined byMTT assay. Apoptosis was detected with Hoeehst 33258 staining and annexin-V FITC/PI double-labeled flowcytometry. The influence on cell cycle was studied by a propidium iodide method. Both flow cytometry(FCM) and RT-PCR techniques were applied to assess the expression of Nup88, whereas the localization ofNup88 was determined by confocal laser scanning microscopy. Results GA presented striking inhibitoryeffect on proliferation of HL-60 cells in vitro and induction of apoptosis in a time- and dose-dependentmanner. However, no obvious influence was found on the cell cycle in HL-60 cells. The IC50 value for 12 hwas 1.797 μmol/L. 15.1% of HL-60 calls went apoptosis when treated with 0.4 μmol/L GA for 12 h.When the dose of GA was increased to 1.6 μmol/L, more than half of cells were apoptotic. On the otherhand, the expression level of Nup88 was down-regulated in HL-60 cells induced by GA in a dose-dependentmanner. The distribution of Nup88 was also changed from widely dispersed in both nucleus and cytoplasm tothat only localized at the cytoplasmic side of nuclear membrane, occasionally in the cytoplasm sporadically.Coneinsion GA exhibites remarkable inhibitory effect on cell proliferation in leukemic cells and inducingapoptosis in HL-60 cells in a cell cycle-independent manner, which might correspond to the regulation of theexpression as well as the distribution of nucleoporin Nup88. It may become a new remedy for treatment foracute leukemia.
