CAJ | 학술논문

目的:研究PI3K/Akt抑制剂渥曼青霉素对白血病细胞增殖和凋亡的影响,并探讨其可能的作用机制.方法:以不同浓度的渥曼青霉素作用于人类髓细胞白血病细胞K562,采用MTT法检测细胞增殖活性,单细胞凝胶电泳技术检测细胞DNA损伤形成的"彗星"样拖尾现象,Annexin V-FITC/PI双标法检测细胞凋亡,Western blotting、RT-PCR检测渥曼青霉素作用K562细胞后总Akt和磷酸化Akt以及NF-κB基因及蛋白表达水平的变化.结果: 渥曼青霉素以时间-剂量依赖性方式抑制K562细胞的增殖,其24 h的IC_(50)是25 nmol/L.渥曼青霉素诱导K562细胞发生凋亡,其作用呈明显剂量依赖性增强.渥曼青霉素作用后K562细胞DNA链断裂,呈现"彗星"拖尾现象,其尾长与拖尾率显著高于对照组(P<0.01).渥曼青霉素能同时在蛋白和基因水平,以剂量依赖性方式抑制磷酸化Akt以及NF-κB表达,但对总Akt蛋白没有明显影响.结论: 渥曼青霉素以时间和剂量依赖方式明显抑制K562细胞的增殖及诱导其凋亡,其机制可能与其下调磷酸化PI3K/Akt信号通路以及NF-κB蛋白表达有关.
목적:연구PI3K/Akt억제제악만청매소대백혈병세포증식화조망적영향,병탐토기가능적작용궤제.방법:이불동농도적악만청매소작용우인류수세포백혈병세포K562,채용MTT법검측세포증식활성,단세포응효전영기술검측세포DNA손상형성적"혜성"양타미현상,Annexin V-FITC/PI쌍표법검측세포조망,Western blotting、RT-PCR검측악만청매소작용K562세포후총Akt화린산화Akt이급NF-κB기인급단백표체수평적변화.결과: 악만청매소이시간-제량의뢰성방식억제K562세포적증식,기24 h적IC_(50)시25 nmol/L.악만청매소유도K562세포발생조망,기작용정명현제량의뢰성증강.악만청매소작용후K562세포DNA련단렬,정현"혜성"타미현상,기미장여타미솔현저고우대조조(P<0.01).악만청매소능동시재단백화기인수평,이제량의뢰성방식억제린산화Akt이급NF-κB표체,단대총Akt단백몰유명현영향.결론: 악만청매소이시간화제량의뢰방식명현억제K562세포적증식급유도기조망,기궤제가능여기하조린산화PI3K/Akt신호통로이급NF-κB단백표체유관.
Objective: To study the effect of wortmannin (WM), a PI3K/Akt inhibitor, on the proliferation and apoptosis of leukemia cells and the possible mechanism. Methods: Human leukemia cell line K562 was treated with different concentrations of WM. The proliferation of K562 cells was examined by MTT assay. DNA damage in K562 cells was examined by single cell gel electrophoresis assay, and apoptosis of K562 cells was detected by Annexin V-FITC/PI double-staining. The expressions of total Akt, phosphorate-Akt (p-Akt), and NF-κB p65 mRNA and protein were detected by RT-PCR and Western blotting, respectively. Results: WM inhibited the proliferation of K562 cells in a dose- and time-dependent manner, with the IC((50) value of 24 h being 25 nmol/L. WM also induced apoptosis of K562 cells in a dose-dependent manner. DNA damage in K562 cells was demonstrated by appearance of comet tail after treatment with WM, with the rate of DNA tail and the tail length being significantly higher than those in the control group (P<0.01). WM dose-dependently inhibited P-Akt and NF-κB p65, but not the total Akt, mRNA and protein expressions. Conclusion: WM can inhibit proliferation and induce apoptosis of K562 cells in a dose- and time-dependent manner, probably through down-regulation of phosphorate PI3K/Akt signal pathway and NF-κB expression.