CAJ | 학술논문

目的：研究XN4在体外抑制急性粒细胞白血病细胞( AML)增殖的作用与诱导DNA损伤的关系。方法 MTT法检测XN4对AML细胞增殖抑制作用；流式细胞术检测AML细胞反应性氧自由基( ROS)、DNA损伤、细胞周期和细胞凋亡；Western blot探讨XN4对相关蛋白表达的影响。结果XN4明显抑制 AML 细胞增殖,半数抑制率( IC50)分别为(2．79±0．15)μmol·L-1和(2．76±0．20)μmol·L-1；XN4增加细胞ROS水平和r-H2AX的表达,诱导细胞阻滞在S期并增加细胞凋亡率；XN4能增加H2AX、ATM的磷酸化以及Parp和 Caspase-3的切割,而减少 CDK2和 Cyclin E1的表达。结论 XN4通过激活ROS,诱导DNA的损伤和细胞周期S期阻滞,抑制 DNA损伤修复和诱导细胞的凋亡,抑制HL-60及KG1α细胞的增殖,抗氧化剂NAC ( N-乙酰半胱氨酸)能减少ROS的产生逆转XN4的作用。
목적：연구XN4재체외억제급성립세포백혈병세포( AML)증식적작용여유도DNA손상적관계。방법 MTT법검측XN4대AML세포증식억제작용；류식세포술검측AML세포반응성양자유기( ROS)、DNA손상、세포주기화세포조망；Western blot탐토XN4대상관단백표체적영향。결과XN4명현억제 AML 세포증식,반수억제솔( IC50)분별위(2．79±0．15)μmol·L-1화(2．76±0．20)μmol·L-1；XN4증가세포ROS수평화r-H2AX적표체,유도세포조체재S기병증가세포조망솔；XN4능증가H2AX、ATM적린산화이급Parp화 Caspase-3적절할,이감소 CDK2화 Cyclin E1적표체。결론 XN4통과격활ROS,유도DNA적손상화세포주기S기조체,억제 DNA손상수복화유도세포적조망,억제HL-60급KG1α세포적증식,항양화제NAC ( N-을선반광안산)능감소ROS적산생역전XN4적작용。
Aim To investigate the cytotoxicity of XN4 against AML cells, and the underlying mechanisms by which XN4 might induce DNA damage and apoptotic cell death through reactive oxygen species ( ROS ) . Methods The proliferation inhibition ratio of AML cells was measured by MTT. The level of extracellular ROS, DNA damage, cell cycle process and apoptosis were tested by flow cytometry ( FCM ) . Western blot was applied to test the expression of proteins. Results XN4 significantly inhibited the proliferation of HL-60 and KG1α with IC50 ( 2. 79 ± 0. 15 ) μmol · L-1 and (2. 76 ± 0. 20) μmol·L-1 respectively. XN4 signifi-cantly increased the generation of intracellular ROS, followed by inducing DNA damage and activating the ATM-γ-H2AX signaling, which led to increases of cells in the S phases of the cell cycle. Subsequently, XN4 induced apoptotic cell death through activation of caspase-3 and Parp. Moreover, the above effects were all reversed by the ROS scavenger N-acetylcysteine ( NAC ) . Conclusion XN4-induced DNA damage and cell apoptosis in AML cells are mediated via ROS generation.