CAJ | 학술논문

目的研究K562细胞株及其耐药细胞株K562/A02 NF-κ B活性的差异性表达,探讨白血病多药耐药(MDR)发病机制.方法用MTT法检测K562/A02的耐药倍数,观察细胞生长的形态学变化,PI单染法检测K562/S、K562/A02细胞周期分布,并用RT-PCR方法榆测mRNA的表达、流式细胞仪检测P糖蛋白(P-gp)表达及其功能,Western blotting方法检测细胞核NF-κB p65表达,比较K562与K562/A02白血病耐药细胞株之间的差异性.结果与K562细胞不同,耐药细胞株K562/A02细胞生长特性发生改变,旱半贴壁生长,与K562/S细胞相比,K562/A02细胞的G0/G1期、S期比例增高,G2/M期细胞比例减低,差异有统计学意义(P＜0.05),凋亡细胞比例差异无统计学意义(P＞0.05),且检测到mdrl mRNA及细胞膜P-gp表达增高以及细胞核内NF-κB p65表达明显增加.结论 NF-κ B信号通路的激活即NF-κ B p65细胞核内转位可能参与了白血病MDR的形成.
목적 연구K562세포주급기내약세포주K562/A02 NF-κ B활성적차이성표체,탐토백혈병다약내약(MDR)발병궤제.방법 용MTT법검측K562/A02적내약배수,관찰세포생장적형태학변화,PI단염법검측K562/S、K562/A02세포주기분포,병용RT-PCR방법유측mRNA적표체、류식세포의검측P당단백(P-gp)표체급기공능,Western blotting방법검측세포핵NF-κB p65표체,비교K562여K562/A02백혈병내약세포주지간적차이성.결과 여K562세포불동,내약세포주K562/A02세포생장특성발생개변,한반첩벽생장,여K562/S세포상비,K562/A02세포적G0/G1기、S기비례증고,G2/M기세포비례감저,차이유통계학의의(P＜0.05),조망세포비례차이무통계학의의(P＞0.05),차검측도mdrl mRNA급세포막P-gp표체증고이급세포핵내NF-κB p65표체명현증가.결론 NF-κ B신호통로적격활즉NF-κ B p65세포핵내전위가능삼여료백혈병MDR적형성.
Objective To explore the different expression of NF-κB in both K562 and its multidrug resistant cell line K562/A02 and discuss the mechanism of muhidrug resistance(MDR). Methods To detect the growing feature of the cells. Flow cytometry was used to analys the difference between the distribution profile of K562/S and K562/A02 cell. MTT colorimetry was used to determine the cytotoxic effect of adramycin, and expression of mdrl gene was detected by semi-quantitative reverse transcriptase poly-merase chain reaction (RT-PCR) in K562 and K562/A02 cells. FACS was used to determine the expression and function of glycoprotein (P-gp) on the cell membrane. Western blotting was used to determine the NF-κB p65protein in nueleus. Results There was a difference between K562 and K562/A02 cells growed in a halfadherent way rather than suspending ones, there were increases in the percentage number of cells at G0/G1 and S phases(P ＜0.05). This was mirrored by a decreasing number of cells within the G2/M phase(P＜0.05). Butthere was no difference in apoptosis rate(P ＞0.05). mdr1 mRNA was detected in K562/A02 cells, in which the expression P-gp was much higher [(94.17±0.89)%:(1.41 ±O.491)%]. NF-κB p65 protein in nucleus was overexpressed in K562/A02 cells. Conclusion The activation of NF-κB signaling pathway may attribute to the formation of MDR in K562/A02 cells.