吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (01): 96-101.doi: 10.13481/j.1671-587x.20200117

• 基础研究 • 上一篇    下一篇

紫草素对人白血病MV4-11细胞的增殖抑制和促凋亡作用

苏龙, 张云蔚, 王卓, 宋飞, 秦田雪, 高素君   

  1. 吉林大学第一医院肿瘤中心血液科, 吉林 长春 130021
  • 收稿日期:2019-10-15 出版日期:2020-01-28 发布日期:2020-02-03
  • 通讯作者: 高素君,教授,博士研究生导师(Tel:0431-88782157,E-mail:drsujung@163.com) E-mail:drsujung@163.com
  • 作者简介:苏龙(1986-),男,内蒙古自治区包头市人,主治医师,医学硕士,主要从事血液病基础和临床方面的研究。
  • 基金资助:
    国家自然科学基金青年基金项目资助课题(81900174);吉林省财政厅人才卫生专项基金资助课题(JLSCZD2019-09)

Proliferation inhibition and pro-apoptotic effects of shikonin on human leukemia MV4-11 cells

SU Long, ZHANG Yunwei, WANG Zhuo, SONG Fei, QIN Tianxue, GAO Sujun   

  1. Department of Hematology, Cancer Center, First Hospital, Jilin University, Changchun 130021, China
  • Received:2019-10-15 Online:2020-01-28 Published:2020-02-03

摘要: 目的:探讨紫草素对FMS样酪氨酸激酶3受体基因内部串联重复序列(FLT3-ITD)突变急性髓系白血病(AML)MV4-11细胞的增殖抑制和促凋亡作用,并初步阐明其分子机制。方法:生长状态良好的MV4-11细胞分为二甲基亚砜(DMSO)组和不同浓度(0.5、1.0、2.0、4.0和8.0 μmol·L-1)紫草素组,处理细胞24和48 h,采用CCK-8法检测各组细胞增殖抑制率,并计算半数抑制浓度(IC50)。MV4-11细胞分为空白对照组、DMSO组和不同浓度(0.25、0.50和1.00 μmol·L-1)紫草素组,处理细胞48和72 h,采用羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记法检测各组细胞增殖率。MV4-11细胞分为DMSO组和不同浓度(0.702、1.404和2.808 μmol·L-1)紫草素组,处理细胞48 h,采用流式细胞术检测各组细胞凋亡率。MV4-11细胞分为DMSO组和不同浓度(0.351、0.702和1.404 μmol·L-1)紫草素组,处理细胞48 h,Real-time PCR法检测各组细胞中微小RNA-155(miR-155)的表达水平。结果:CCK-8法和CFSE法检测,与DMSO组比较,不同浓度紫草素组细胞增殖抑制率明显升高(P<0.05或P<0.01),增殖率明显降低(P<0.05),且呈剂量依赖性;24和48 h时IC50分别为1.743和1.404 μmol·L-1。流式细胞术检测,与DMSO组比较,不同浓度紫草素组细胞凋亡率明显升高(P<0.01),且呈剂量依赖性。Real-time PCR法检测,与DMSO组比较,不同浓度紫草素组细胞中miR-155表达水平明显降低(P<0.01),1.404 μmol·L-1紫草素组细胞中miR-155表达水平降低超过75%。结论:紫草素对人FLT3-ITD突变AML MV4-11细胞具有增殖抑制和促凋亡作用,并可下调miR-155表达,提示紫草素可能作为FLT3-ITD突变AML的潜在治疗药物之一。

关键词: 急性髓系白血病, FMS样酪氨酸激酶3受体基因内部串联重复序列, 基因突变, 紫草素, 微小RNA-155

Abstract: Objective: To discuss the proliferation inhibition and apoptosis induction of shikonin on the FMS-like tyrosine kinase-3 receptor internal tandem duplication (FLT3-ITD) mutated acute myeloid leukemia (AML) MV4-11 cells, and to preliminarily clarify the molecular mechanisms. Methods: The MV4-11 cells were divided into DMSO group and different concentrations (0.5, 1.0, 2.0, 4.0, and 8.0 μmol·L-1) of shikonin groups, and treated for 24 and 48 h. The inhibitory rate of proliferation was analyzed by CCK-8 assay, and half inhibitory concentration (IC50) was calculated. The MV4-11 cells were divided into blank control group, DMSO group, and different concentrations (0.25, 0.50, and 1.00 μmol·L-1) of shikonin groups, and treated for 48 and 72 h; the proliferation rate of cells was analyzed by carbox fluorescenceindiacetate succinimidyl este (CFSE). The MV4-11 cells were divided into DMSO group and different concentrations (0.702, 1.404, and 2.808 μmol·L-1) of shikonin groups, and treated for 48 h;the apoptotic rate was determined by flow cytometry. The MV4-11 cells were divided into DMSO group and different concentrations (0.351, 0.702, and 1.404 μmol·L-1) of shikonin groups, and treated for 48 h; the microRNA-155 (miR-155) expression level was detected by Real-time PCR. Results: The results of CCK-8 and CFSE methods indicated that the inhibitory rates of proliferation of MV4-11 cells in different concentrations of shikonin groups were increased compared with DMSO grpup(P<0.05 or P<0.01), and the proliferation rates were decreased (P<0.05 or P<0.01) in a concentration-dependent manner; the IC50 of 24 and 48 h were 1.743 and 1.404 μmol·L-1, respectively. The flow cytometry results showed that the apoptotic rates of the cells in different concentrations of shikonin groups were increased compared with DMSO group (P<0.01) in a concentration-dependent manner. The Real-time PCR results showed that the expression levels of miR-155 in the cells in different concentrations of shikonin groups were decreased significantly(P<0.01), and the expression level in 1.404 μmol·L-1 shikonin group was decreased by more than 75%. Conclusion: Shikonin could inhibit the proliferation and promote the apoptosis of FLT3-ITD mutated AML MV4-11 cells, and down-regulate the expression of miR-155, suggesting that shikonin may be one of the potential therapeutic drugs for FLT3-ITD mututed AML.

Key words: acute myeloid leukemia, FMS-like tyrosine kinase-3 receptorinternal tandem duplication, mutation, shikonin, microRNA-155

中图分类号: 

  • R73-361