CAJ | 학술논문

目的探讨2-甲氧基雌二醇(2-ME)对白血病K562细胞凋亡的影响及其作用机制.方法实验分为3组,对照组:培养基中不含2-ME;实验组:分别用1、2.4、8、16μmol/L的2-ME处理K562细胞36 h后观察各项指标的变化;阴性对照组:培养液中以无RNase蒸馏水代替K562细胞.应用原位细胞凋亡法(TUNEL)、流式细胞术(FCM)、半定量RT-PCR及黄嘌呤氧化酶法分别检测K562细胞凋亡率、Caspase-3、性连锁凋亡抑制蛋白(XIAP)及其基因表达、超氧化物歧化酶(SOD)与活性氧(ROS)水平.结果不同浓度的2-ME与K562细胞作用36 h后,2-ME在一定的浓度范围内以剂量依赖形式诱导K562细胞凋亡;2-ME通过上调促凋亡因子Caspase-3和(或)下调抗凋亡因子XIAP的表达,降低SOD及增加ROS水平等途径促进K562细胞凋亡,这可能是2-ME诱导K562细胞凋亡的机制之一.结论 2-ME能够以剂量依赖形式诱导K562细胞凋亡,显示了其在慢性粒细胞白血病(CML)治疗中的潜在价值.
목적 탐토2-갑양기자이순(2-ME)대백혈병K562세포조망적영향급기작용궤제.방법 실험분위3조,대조조:배양기중불함2-ME;실험조:분별용1、2.4、8、16μmol/L적2-ME처리K562세포36 h후관찰각항지표적변화;음성대조조:배양액중이무RNase증류수대체K562세포.응용원위세포조망법(TUNEL)、류식세포술(FCM)、반정량RT-PCR급황표령양화매법분별검측K562세포조망솔、Caspase-3、성련쇄조망억제단백(XIAP)급기기인표체、초양화물기화매(SOD)여활성양(ROS)수평.결과 불동농도적2-ME여K562세포작용36 h후,2-ME재일정적농도범위내이제량의뢰형식유도K562세포조망;2-ME통과상조촉조망인자Caspase-3화(혹)하조항조망인자XIAP적표체,강저SOD급증가ROS수평등도경촉진K562세포조망,저가능시2-ME유도K562세포조망적궤제지일.결론 2-ME능구이제량의뢰형식유도K562세포조망,현시료기재만성립세포백혈병(CML)치료중적잠재개치.
Objective To investigate the effects of 2-methoxyestradiol( 2-ME) on apoptosis of K562 cells and its mechanisms. Methods The K562 cells were cultured and divided into three groups. The control group: K562 cells were cultured without 2-ME treatment. The experimental group: K562 cells were cultured in the medium containing different concentrations of 2-ME (1, 2, 4, 8 and 16 μmol/L) for 36 h. The negative control group: K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36 h. The apoptosis rate, the protein and its mRNA expression of Caspase-3 and XIAP, the activity of superoxide dismutase (SOD) and reactive oxygen species (ROS) of K562 cells wasdetected by TUNEL, flow cytometry (FCM), half-quantitative RT-PCR and xanthenes oxidized enzyme assay,respectively. Results After treated with 2-ME at different concentrations for 36 hours, in the specified concentration range, 2-ME induced apoptosis of K562 cells in a concentration-dependent manner. The possible functional mechanism of 2-ME was to up-regulate Caspase-3 but down-regulate XIAP mRNA expression, and increase ROS activity but decrease SOD activity. Conclusion 2-ME can induce apoptosis of K562 cells in a concentration-dependent manner and indicate its promising potential in the treatment of chronic myeloid leukemia(CML) patients.