Abstract：
Chronic Myeloid Leukaemia (CML) starts in certain blood-forming cells of the bone marrow when cells acquire Philadelphia chromosome. Nowadays, scientists attempt to find novel and safe therapeutic agents and approaches for CML therapy using Tyrosine Kinase Inhibitors (TKIs), CML conventional treatment agents, has some restrictions and also adverse effects. Recently, it has been proposed that phytochemicals, such as flavonoids due to their low side effects and notable safety have the potential to be used for CML therapy.
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.
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.
摘要：
当细胞获得费城染色体时，慢性髓系白血病（CML）在骨髓的某些造血细胞中开始。如今,科学家们试图寻找新的和安全的治疗剂和使用酪氨酸激酶抑制剂（TKIs）治疗慢性粒细胞白血病的方法，CML常规治疗剂,有一些限制，也有不利影响。最近,有人提出植物化学物质，类黄酮由于其副作用低和显著的安全性，有可能用于CML治疗。
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。
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。
