从山香圆中提取的一种多甲氧基黄酮可通过激活内在凋亡途径诱导伊马替尼耐药 K562R 细胞凋亡。

A polymethoxyflavone from Laggera pterodonta induces apoptosis in imatinib-resistant K562R cells via activation of the intrinsic apoptosis pathway.

机构信息

Department of Human Anatomy, Medical School of Jinan University, Guangzhou, 510632 China ; Institute of Hematology, Jinan University, Guangzhou, 510632 China.

Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, Jinan University, Guangzhou, China.

出版信息

Cancer Cell Int. 2014 Dec 5;14(1):137. doi: 10.1186/s12935-014-0137-1. eCollection 2014.

DOI:10.1186/s12935-014-0137-1

PMID:25530716

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4272561/

Abstract

BACKGROUND

Treatment with imatinib mesylate (IM) (a tyrosine kinase inhibitor) is the first line of standard care for patients newly diagnosed with CML. Despite the success of IM and other tyrosine kinase inhibitors (TKIs), chronic myeloid leukemia (CML) remains largely incurable, and a number of CML patients die due to Abl mutation-related drug resistance and blast crisis. 3, 5-Dihydroxy-6, 7, 3'4'-tetramethoxyflavone (DHTMF) is a polymethoxyflavone isolated from Laggera pterodonta which is a herbal medicine used to treat cancer in the Chinese folk. In the previous study, we found DHTMF demonstrated good antiproliferative activities against a number of cancer cell lines and induced the apoptosis of CNE cells in vitro in a time- and dose-dependent manner while exhibiting low cytotoxicity in the two normal cell lines Vero and EVC304. The aim of the present study was to evaluate the proliferation inhibition and apoptosis induced by DHTMF alone and in combination with IM in the IM-resistant CML cell line K562R.

METHODS

Cell proliferation was assayed with the cell counting kit-8 (CCK8) method. The apoptosis percentage was determined by flow cytometry (FCM). Mitochondrial transmembrane potential was detected using FCM and confocal laser-scanning microscopy. The level of proteins involved in apoptosis was detected by Western blotting.

RESULTS

DHTMF suppressed K562R cell viability in both time- and dose-dependent manners. DHTMF combined with IM enhanced the inhibitory effects and apoptosis in K562R cells as compared with DHTMF alone. DHTMF alone and in combination with IM significantly decreased the mitochondrial membrane potential and increased the levels of cleaved caspase-9, caspase-7, caspase-3, and PARP in K562R cells.

CONCLUSIONS

We demonstrated that DHTMF could inhibit IM-resistant K562R cell proliferation and induces apoptosis via the intrinsic mitochondrial apoptotic pathway. These results suggest that DHTMF may be a potential therapeutic drug with lower side effects against IM resistance in CML cells.

摘要

背景

伊马替尼甲磺酸盐（IM）（一种酪氨酸激酶抑制剂）的治疗是新诊断为 CML 的患者的一线标准治疗方法。尽管 IM 和其他酪氨酸激酶抑制剂（TKI）取得了成功，但慢性髓性白血病（CML）仍然在很大程度上无法治愈，许多 CML 患者因 Abl 突变相关耐药性和急变期而死亡。3,5-二羟基-6,7,3',4'-四甲氧基黄酮（DHTMF）是从拉格gera pterodonta 中分离出来的多甲氧基黄酮，是一种民间草药，用于治疗癌症。在之前的研究中，我们发现 DHTMF 对多种癌细胞系表现出良好的抗增殖活性，并在体外以时间和剂量依赖的方式诱导 CNE 细胞凋亡，同时在两个正常细胞系 Vero 和 EVC304 中表现出低细胞毒性。本研究旨在评估 DHTMF 单独和与 IM 联合在 IM 耐药 CML 细胞系 K562R 中的增殖抑制和凋亡诱导作用。

方法

用细胞计数试剂盒-8（CCK8）法测定细胞增殖。通过流式细胞术（FCM）测定凋亡百分比。用 FCM 和共聚焦激光扫描显微镜检测线粒体跨膜电位。用 Western blot 检测参与凋亡的蛋白水平。

结果

DHTMF 以时间和剂量依赖的方式抑制 K562R 细胞活力。DHTMF 与 IM 联合使用可增强 DHTMF 对 K562R 细胞的抑制作用和凋亡作用。DHTMF 单独和与 IM 联合使用可显著降低线粒体膜电位并增加 K562R 细胞中 cleaved caspase-9、caspase-7、caspase-3 和 PARP 的水平。