大黄素甲醚对白血病耐药细胞系K562/ADM耐药性的影响及机制研究

Reverse effect of Physcion on multi-drug resistance of leukemic drug-resistant cell line K562/ADM

摘要:

目的 评价大黄素甲醚对慢性粒细胞白血病（CML）耐药细胞系K562/ADM 多药耐药的逆转作 用及其机制。方法 CCK-8、克隆形成实验检测细胞的增殖变化，流式细胞仪、Hoechst 染色检测细胞的凋亡， 实时荧光定量聚合酶链反应（qRT-PCR）、Western blot 检测各相关基因的表达，迁移实验检测细胞的侵袭 能力。结果 大黄素甲醚能增强K562/ADM 细胞对ADM 的敏感性，耐药逆转倍数在2 和5 μmol/L 的浓度下 分别为2.03 和5.30 倍。miRNA-146a 在K562 耐药细胞系中低于K562 细胞，而其在K562/ADM 细胞中过 表达能恢复细胞对ADM 的敏感性。大黄素甲醚可以通过诱导miRNA-146a 的表达抑制CXCL12/CXCR4 信 号通路从而能增强ADM 抗肿瘤细胞增殖的作用。结论 大黄素甲醚可以通过上调miRNA-146a 的表达抑制 CXCL12/CXCR4 信号从而逆转K562/ADM 细胞对ADM 的耐药性。

Abstract:

Objective To investigate the reverse effect of Physcion on multi-drug resistance of chronic myeloid leukemia (CML) cell line K562 / ADM. Methods CCK8 and clone formation assay were used to detect cellular proliferation. Flow cytometry and Hoechst 33258 were performed to determine apoptosis. Expression of related genes was measured by qRT-PCR and Western blot. Migration assay was identified by cell invasiveness assay. Results Physcion increased the sensitivity of K562 / ADM cells to ADM significantly. Fold change of sensitivity induced by Physcion was 2.03 fold and 5.3 fold at concentration of 2 and 5 μmol/L, respectively. miRNA-146a in K562 / ADM cells decreased significantly when compared with K562 cells, and overexpression of miRNA-146a increased the sensitivity to ADM. Physcion-induced upregulated expression of miRNA-146a facilitated inhibition of CXCL12 / CXCR4 signaling pathway. Conclusions Physcion reverses the multi-drug resistance of K562/ADM cells through modulating miRNA-146a mediated CXCL12 / CXCR4 signaling pathway.