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槲皮素对多发性骨髓瘤细胞增殖和凋亡的影响及其相关机制

[Effect of Quercetin on Proliferation and Apoptosis of Multiple Myeloma Cells and Its Related Mechanism].

作者信息

Xu Ya-Wen, Zou Li-Fang, Li Fei

机构信息

Department of Hematology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China,Institute of Hematology, Academy of Clinical Medicine of Jiangxi Province, Nanchang 330006, Jiangxi Province, China.

Department of Hematology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China,Institute of Hematology, Academy of Clinical Medicine of Jiangxi Province, Nanchang 330006, Jiangxi Province, China,E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Aug;28(4):1234-1239. doi: 10.19746/j.cnki.issn.1009-2137.2020.04.025.

Abstract

OBJECTIVE

To investigate the effect of quercetin (que) on proliferation and apoptosis of multiple myeloma cell line NCI-H929.

METHODS

NCI-H929 cells were routinely cultured, and cells in logarithmic growth phase were taken and used for experiments. After treatment of NCI-H929 cells with Que of 50, 100 and 200 µmol/ L for 24, 48 and 72 hours, the proliferation level of cells was detected by using MTT method; after treatment of NCI-H929 cells with Que of 100 and 200 µmol/ L for 24 hours, the cell apoptosis level was detected by Annexin V-FITC/PI double staining, the changes of cell cycle was analysis by flow cytometry with PI marking; the expression of apoptosis-related proteins caspase-3, caspase-8, caspase-9, PARP, BCL-2 and cell cycle-related proteins P53, P21, P27, CDK4, and the activiation of ERK and ATK were detected by Western blot.

RESULTS

Que of different concentration could inhibit cell proliferation with time and dose dependent manner. The flow cytometry showed that Que could significantly increase the cell apoptosis and arrest NCI-H929 cells in the G/M phase. In addition, Western blot analysis showed that Que could activate the apoptosis-related proteins, such as caspase-3, caspase-8, caspase-9 and PARP, and then inhibited the expression of BCL-2. Que could promote the expression of P53, P21 and P27, however, it could inhibited the CDK4 expression in NCI-H929 cells. Que could decrease the phosphorylation levels of p-ERK and p-AKT in NCI-H929 cells.

CONCLUSION

Quercetin mediates anti-myeloma effects through inducing apoptosis, cell cycle arrest and down-regulating ERK and AKT pathways in human myeloma cells.

摘要

目的

探讨槲皮素(que)对多发性骨髓瘤细胞系NCI-H929增殖和凋亡的影响。

方法

常规培养NCI-H929细胞,取对数生长期细胞用于实验。用50、100和200μmol/L的Que处理NCI-H929细胞24、48和72小时后,采用MTT法检测细胞增殖水平;用100和200μmol/L的Que处理NCI-H929细胞24小时后,采用Annexin V-FITC/PI双染法检测细胞凋亡水平,用PI标记通过流式细胞术分析细胞周期变化;采用蛋白质免疫印迹法检测凋亡相关蛋白caspase-3、caspase-8、caspase-9、PARP、BCL-2以及细胞周期相关蛋白P53、P21、P27、CDK4的表达,以及ERK和ATK的激活情况。

结果

不同浓度的Que可抑制细胞增殖,呈时间和剂量依赖性。流式细胞术显示,Que可显著增加细胞凋亡,并使NCI-H929细胞停滞于G/M期。此外,蛋白质免疫印迹分析显示,Que可激活凋亡相关蛋白,如caspase-3、caspase-8、caspase-9和PARP,进而抑制BCL-2的表达。Que可促进P53、P21和P27的表达,然而,它可抑制NCI-H929细胞中CDK4的表达。Que可降低NCI-H929细胞中p-ERK和p-AKT的磷酸化水平。

结论

槲皮素通过诱导人骨髓瘤细胞凋亡、细胞周期停滞以及下调ERK和AKT信号通路发挥抗骨髓瘤作用。

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