植物鞘氨醇诱导白血病细胞凋亡的作用及机制研究

杨官翠; 刘金宜; 蒋佩洁; 许玉溪; 田小龙; 王筱淇; 王瑞; 杨世杰; 宋清晓; 魏锦; 张曦

doi:10.16016/j.2097-0927.202310010

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植物鞘氨醇诱导白血病细胞凋亡的作用及机制研究

专题报道：血液肿瘤诊治前沿 | 更新时间：2026-01-09

- 植物鞘氨醇诱导白血病细胞凋亡的作用及机制研究
- Anti-tumor effects of phytosphingosine on leukemia cells by inducing cell apoptosis
- 陆军军医大学学报 2024年46卷第4期页码：359-368
- 作者机构：
  
  1.637002 南充，川北医学院附属医院血液内科
  2.400037 重庆，陆军军医大学(第三军医大学)第二附属医院血液病医学中心，全军临床重点专科，创伤与化学中毒全国重点实验室，重庆市临床重点专科，血液病与微环境重庆市重点实验室
  3.401329 重庆，金凤实验室
- 作者简介：
  
  [魏锦，E-mail: 1497058875@qq.comWEI Jin, E-mail: 1497058875@qq.com
  [张曦，E-mail: zhangxxi@sina.comZHANG Xi, E-mail: zhangxxi@sina.com
- 基金信息：
  
  国家自然科学基金青年科学基金项目(82200242);重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX1060);重庆市自然科学基金面上项目(CSTB2023NSCQ-MSX0262);新桥医院青年博士孵育计划项目(2022YQB062);作者贡献声明杨官翠、刘金宜参与本研究的实验方案设计、实验操作、数据整理及分析、实验结果统计、初稿撰写及修改；蒋佩洁、许玉溪、田小龙参与本研究实验操作、数据整理及分析、实验结果统计；王筱淇、王瑞、杨世杰、宋清晓参与实验结果可视化、文章的修改；魏锦、张曦负责本研究的设计与构想、结果的审核、实验的监督与管理、稿件审阅与修改等
- DOI：10.16016/j.2097-0927.202310010
  中图分类号： R733.7;R965;R977.6
- 收稿：2023-10-06，
  
  修回：2023-12-22，
  
  纸质出版：2024-02-28
- 稿件说明：
移动端阅览
杨官翠, 刘金宜, 蒋佩洁, 等. 植物鞘氨醇诱导白血病细胞凋亡的作用及机制研究[J]. 陆军军医大学学报, 2024,46(4):359-368.

Guancui YANG, Jinyi LIU, Peijie JIANG, et al. Anti-tumor effects of phytosphingosine on leukemia cells by inducing cell apoptosis[J]. Journal of Army Medical University, 2024, 46(4): 359-368.
杨官翠, 刘金宜, 蒋佩洁, 等. 植物鞘氨醇诱导白血病细胞凋亡的作用及机制研究[J]. 陆军军医大学学报, 2024,46(4):359-368. DOI： 10.16016/j.2097-0927.202310010.

Guancui YANG, Jinyi LIU, Peijie JIANG, et al. Anti-tumor effects of phytosphingosine on leukemia cells by inducing cell apoptosis[J]. Journal of Army Medical University, 2024, 46(4): 359-368. DOI： 10.16016/j.2097-0927.202310010.

摘要

目的

初步探讨植物鞘氨醇(phytosphingosine

PHS)对白血病细胞凋亡的影响及其可能机制。

方法

采用人白血病细胞株K562及SUP-B15作为研究模型

CCK-8法检测细胞活力

EdU检测细胞DNA合成能力

RNA-seq检测PHS处理后差异表达基因

GO功能富集及KEGG信号通路富集分析PHS可能作用的分子途径

CTD数据库和Discovery Studio软件对PHS进行反向找靶

Discovery Studio软件对PHS和BCL-2进行分子对接

流式细胞术检测细胞凋亡变化

JC-1染色检测线粒体膜电位变化

Western blot检测凋亡相关分子蛋白水平表达变化。

结果

PHS能抑制K562和SUP-B15细胞增殖

且呈剂量和时间依赖性。PHS的半数抑制浓度在K562细胞中分别为17.67 μmol/L (24 h)和12.52 μmol/L (48 h)

在SUP-B15细胞中分别为17.58 μmol/L (24 h)和14.86 μmol/L (48 h)。20 μmol/L PHS作用白血病细胞48 h后

能显著抑制细胞DNA的合成(

0.05)。对PHS作用于K562细胞后差异基因进行GO富集分析发现PHS可能参与细胞凋亡正向调控、细胞质膜及组分、激酶结合与活性等生物功能。反向找靶结果提示

BCL-2作为PHS作用靶点的可能性最高。PHS作用白血病细胞48 h后细胞凋亡率显著增加(

0.05)

呈剂量依赖性; 线粒体膜电位下降

抗凋亡蛋白BCL-2表达下调(

0.05)

Cleaved caspase-3及Cleaved caspase-9蛋白表达上调(

0.05)。

结论

PHS诱导白血病细胞发生线粒体依赖性细胞凋亡

抑制白血病细胞增殖

并可能与PHS/BCL-2结合体有关。

Abstract

Objective

To preliminarily investigate the anti-tumor effects of phytosphingosine (PHS) and the involvement of inducing apoptosis of leukemia cells.

Methods

Cellular model of leukemia was established in leukemia cell lines K562 and SUP-B15.CCK-8 assay and EdU assay were used to measure the viability and DNA synthesis of K562 and SUP-B15 cells.RNA-seq was carried out to verify the differentially expressed genes (DEGs) after PHS treatment.Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were applied to analyze the involved functions and signaling pathways.Comparative Toxicogenomics Database (CTD) and Discovery Studio software were employed to predict the underlying targets of PHS and molecular docking.Cell apoptosis was detected by flow cytometry

mitochondrial membrane potential was evaluated by JC-1 probe

and protein expression of key molecules was validated by Western blotting.

Results

PHS inhibited the proliferation of K562 and SUP-B15 cells in a time-and dose-dependent manner.The half-maximal inhibitory concentration (IC

) of K562 cells was 17.67 and 12.52 μmol/L for 24 and 48 h

respectively

and the IC

value of SUP-B15 cells was 17.58 and 14.86 μmol/L for 24 and 48 h

respectively.PHS treatment at a dose of 20 μmol/L for 48 h resulted in significant inhibition of DNA synthesis.GO enrichment analysis of the K562 cells showed that PHS might be involved in positive regulation of apoptotic process

plasma membrane and its integral components

and protein kinase binding and activity.Reverse predictive analysis showed that BCL-2 protein was the most likely target of PHS.PHS significantly increased the apoptotic rate of leukemia cells (

0.05) in a dose-dependent manner

reduced the mitochondrial membrane potential

and down-regulated BCL-2 level (

0.05) and up-regulated the levels of Cleaved caspase-3 and Cleaved caspase-9(

0.05).

Conclusion

PHS may inhibit the proliferation of leukemia cells by inducing mitochondria-dependent apoptosis

possibly through PHS and BCL-2 interaction.

关键词

Keywords

references

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