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

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5μmol/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。

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 5μmol/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.