汉防己甲素对白血病骨髓间充质干细胞相关耐药的影响及其机制研究
Study on the Effect and Mechanism of Tetrandrine on Bone Marrow Mesenchymal Stem Cell-Mediated Drug Resistance in Leukemia
周芯夷 1金楠 2陈宝安2
作者信息
- 1. 东南大学医学院
- 2. 东南大学附属中大医院血液科,江苏南京 210009
- 折叠
摘要
目的:研究骨髓间充质干细胞(BMSC)是否对白血病的多药耐药有促进作用,并研究汉防己甲素(TET)对BMSC介导的多药耐药(MDR)的影响及其作用机制.方法:构建人骨髓间充质干细胞与白血病K562细胞株的混合共培养体系和Transwell小室共培养体系,并将细胞进行分组,分别给以柔红霉素(DNR)单药干预、TET与DNR联合干预,然后利用CCK-8法检测各组细胞增殖情况,计算细胞抑制率;利用流式细胞术测定各组上清液中IFN、IL-2、IL-6、IL-10的表达水平;利用RT-qPCR和Western blot分别在mRNA和蛋白水平对K562细胞中STAT3的表达进行验证.结果:与BMSC共培养后,DNR对K562细胞增殖的抑制率明显降低(P＜0.05),培养上清液中IL-6的水平和K562细胞中磷酸化的STAT3水平显著增加(P＜0.05).加入TET联合干预后,DNR对K562细胞增殖的抑制率显著增加(P＜0.05),IL-6和磷酸化的STAT3水平降低(P＜0.05).结论:BMSC对白血病细胞的耐药有促进作用,TET可能通过抑制IL-6/STAT3通路从而逆转骨髓微环境介导的耐药作用.
Abstract
Objective:To explore the role of bone marrow mesenchymal stem cells(BMSC),an essential element of the bone marrow microenvironment,in multidrug resistance(MDR)of K562 cells,as well as the reversal effect of tetrandrine(TET)on BMSC-mediated MDR and its potential mechanism.Methods:A mixed co-culture system and a transwell co-culture system for BMSC and K562 cells were established,and the cells were divided into different groups and treated with daunorubicin(DNR)alone or combined with TET and DNR.The CCK-8 assay was used to detect the proliferation of K562 cells in each group,and the cell inhibition rate was calculated.Cytometric bead array(CBA)was used to detect the expression levels of IFN,IL-2,IL-6 and IL-10 in the supernatant of different groups.RT-qPCR and Western blot were used to detected the expression of STAT3 at mRNA and protein levels,respectively.Results:Compared with K562+DNR group,the inhibition rate of DNR on K562 cell proliferation in K562+BMSC+DNR group was significantly decreased(P＜0.05),while the levels of IL-6 in the culture supernatant and phosphorylated STAT3 in K562 cells were significantly increased(P＜0.05).Compared with K562+BMSC+DNR group,the inhibition rate of DNR on K562 cell proliferation in K562+BMSC+DNR+TET group was significantly increased(P＜0.05),while the level of IL-6 and phosphorylated STAT3 was significantly decreased(P＜0.05).Conclusion:BMSC can promote the drug resistance of leukemia cells,and TET may reverse the BMSC-mediated drug resistance via inhibiting IL-6/STAT3 signaling pathway.
关键词
汉防己甲素/白血病/骨髓间充质干细胞/多药耐药/STAT3Key words
tetrandrine/leukemia/bone marrow mesenchymal stem cells/multidrug resistance/STAT3引用本文复制引用
基金项目
江苏省医学重点学科建设项目(ZDXKB201620)
南京市卫生科技发展项目(YKK22278)
江苏省"六大人才高峰"高层次人才项目(WSE-033)
江苏省青年医学人才项目(QNRC20160813)
出版年
2024
NETL
NSTL
维普
万方数据