K-562 cells were exposed with three concentrations of the querectin (10, 40 and 80µM) for 12, 24 and 48 hours. After that, these cells apoptosis rate was estimated using Annexin-V/PI staining and flowcytometry analysis, and their proliferation rate was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT). Finally, the expression of the 70 and 90 kilodalton heat shock proteins (HSP70 and 90), methionine adenosyltransferase 2A (MAT2A), Forkhead box protein M1 (FOXM1), caspase-3 and -8, Bcl-X(L) and Bax involved in leukemic cells survival and proliferation was assessed using Real-Time PCR within 12, 24 and 48 hours after exposure with quercetin 40 and 80µM.
Considering consequences, querecetin induced apoptosis in K-562 cells, and also abrogated these cells proliferation. On the other hand, RT-PCR results showed a reduction in some of the candidate genes expression, especially HSP70, Bcl-X(L) and FOXM1, when cells were treated with quercetin 40 and 80µM. Also, Bax, caspase-3 and caspase-8 expression was significantly improved in K-562 cells upon quercetin exposure.
We concluded that CML therapy by querecetin due to its anti-proliferative and anti-survival potentials could lead to the promising therapeutic outcome through targeting major survival and proliferation involved genes expression.
将K-562细胞暴露于三种浓度的四肽素(10、40和80µM)12、24和48小时。之后,使用膜联蛋白V/PI染色和流式细胞术分析估计这些细胞的凋亡率,并使用3-[4,5-二甲基噻唑-2-基]-2,5二苯基四唑溴化物(MTT)评估其增殖率。最后,70和90千克热休克蛋白(HSP70和90)的表达,甲硫氨酸腺苷转移酶2A(MAT2A),叉头盒蛋白M1(FOXM1),caspase-3和-8,Bcl-X(L)和Bax与白血病细胞存活和增殖有关,在暴露于槲皮素40和80µM后12、24和48小时内使用Real-TimePCR评估。
考虑到后果,槲皮素诱导K-562细胞凋亡,并废除了这些细胞的增殖。另一方面,RT-PCR结果显示一些候选基因的表达减少,特别是HSP70,Bcl-X(L)和FOXM1,当用槲皮素40和80µM处理细胞时。此外,Bax,槲皮素暴露后,K-562细胞中caspase-3和caspase-8的表达显着提高。
我们得出的结论是,由于其抗增殖和抗存活的潜力,通过靶向主要存活和增殖涉及的基因表达,喹erietin治疗CML可能导致有希望的治疗结果。