Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics

Qing-fang ZENG; Cai-rong LUO; Xiao-yan ZHANG; Yan LIANG; Xiang-chun SHEN; Ling TAO; Wei ZHOU

doi:10.13422/j.cnki.syfjx.20201161

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Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics

Drug Metabolism | 更新时间：2020-12-10

- Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 21, Pages: 173-180(2020)
- 作者机构：
  
  贵州医科大学药学院，天然药物资源优效利用重点实验室，贵阳 550025
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20201161
  CLC： R22;Q51;R28;Q2;R96
- Received：07 January 2020，
  
  Published Online：26 March 2020，
  
  Published：05 November 2020
- 稿件说明：
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曾庆芳,罗才荣,张晓燕等.基于蛋白质组学分析色胺酮抑制小鼠乳腺癌的作用机制[J].中国实验方剂学杂志,2020,26(21):173-180.

ZENG Qing-fang,LUO Cai-rong,ZHANG Xiao-yan,et al.Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):173-180.
曾庆芳,罗才荣,张晓燕等.基于蛋白质组学分析色胺酮抑制小鼠乳腺癌的作用机制[J].中国实验方剂学杂志,2020,26(21):173-180. DOI： 10.13422/j.cnki.syfjx.20201161.

ZENG Qing-fang,LUO Cai-rong,ZHANG Xiao-yan,et al.Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):173-180. DOI： 10.13422/j.cnki.syfjx.20201161.

摘要

目的

采用非标记定量（Label-free）蛋白质组学技术研究色胺酮抗小鼠体内乳腺癌的作用机制。

方法

采用超高效液相色谱-质谱联用技术检测色胺酮抗小鼠乳腺癌的表达蛋白，选择Ionoptics nano UPLC C

色谱柱（0.075 mm×250 mm，1.6 μm），流动相0.1%甲酸水溶液-0.1%甲酸乙腈溶液梯度洗脱，正离子模式，扫描范围

100~1 700，使用MaxQuant 1.6.5.0进行数据库检索。采用Label-free高分辨质谱的蛋白质组学技术筛选4T1乳腺癌小鼠模型组与色胺酮（100 mg·kg

-1

）口服给药组之间的差异表达蛋白，进行色胺酮抗乳腺癌的蛋白质组学研究。

结果

共鉴定出3 997个蛋白质，其中有2 911个蛋白可定量。模型组与色胺酮组共750个差异表达蛋白，其中286个蛋白上调，464个蛋白下调。基因本体分析表明，这些差异表达蛋白主要参与增殖、细胞迁移、凋亡、免疫、血管生成和炎症调节等生物学过程。京都基因与基因组百科全书通路分析进一步表明，这些蛋白主要集中于T细胞受体，B细胞受体，Toll样受体，核转录因子-

B（NF-

B），Ras蛋白，白细胞介素-17，肿瘤坏死因子，磷脂酰肌醇3-激酶/蛋白激酶B（PI3K-Akt）和丝裂原活化蛋白激酶（MAPK）等信号通路。

结论

与色胺酮抗4T1乳腺癌作用密切相关的差异表达蛋白包括上调蛋白白细胞分化抗原14（CD14），前列腺素G/H合酶2（PTGS2），泛素蛋白连接酶E3和下调蛋白CD44，70 kDa热休克蛋白1A（HSPA1A），巨噬细胞移动抑制因子（MIF），NF-

B，核糖体蛋白S6激酶

-4（RPS6KA4）和高迁移率族蛋白B1（HMGB1），提示色胺酮主要通过调节肿瘤炎症微环境来达到抑制小鼠乳腺癌的作用。

Abstract

Objective

A systematical study on the anti-breast cancer mechanism of tryptanthrin in breast cancer-bearing mice was done by Label-free proteomics.

Method

UPLC-MS was used to detect the expressed-proteins of tryptanthrin inhibiting breast cancer in mice， chromatographic separation was achieved on the Ionoptics nano UPLC C

column （0.075 mm×250 mm， 1.6 μm）， and gradient elution was performed with 0.1% formic acid aqueous solution-0.1% formic acid acetonitrile solution as mobile phase. Data acquisition was carried out in electrospray ionization （ESI） under the positive ion mode， the scanning range was

100-1 700， MaxQuant 1.6.5.0 was used for database retrieval. Label-free proteomics with high resolution mass spectrometry was used to screen differentially expressed proteins between the model group of 4T1 breast cancer mice and oral administration group of tryptanthrin （100 mg·kg

-1

）. The proteomics of tryptanthrin against breast cancer was carried out.

Result

A total of 3 997 proteins were identified in this proteomics research， and 2 911 proteins were quantifiable. A total of 750 differentially expressed proteins were identified between the model group and the tryptanthrin group， 286 proteins were up-regulated and 464 proteins were down-regulated. Gene ontology analysis showed that these differentially expressed proteins were mainly involved in biological processes of proliferation， cell migration， apoptosis， immunity， angiogenesis， inflammatory regulation， etc. Kyoto encyclopedia of genes and genomes pathway analysis further indicated that these proteins were mainly concentrated in T cell receptors， B cell receptors， Toll-like receptors， nuclear transcription factor-

B （NF-

B）， Ras proteins， interleukin-17， tumor necrosis factor， phosphatidylinositol 3-kinase/protein kinase B （PI3K-Akt）， mitogen-activated protein kinase （MAPK） and other signaling pathways.

Conclusion

The differentially expressed proteins closely related to anti-breast cancer effect of tryptanthrin on 4T1 breast cancer mice are effectively screened out， including up-regulating proteins of leukocyte differentiation antigen 14 （CD14）， prostaglandin G/H synthase 2 （PTGS2）， E3 ubiquitin-protein ligase and down-regulating proteins of CD44， heat shock 70 kDa protein 1A （HSPA1A）， macrophage migration inhibitory factor （MIF）， NF-

B， ribosomal protein S6 kinase alpha-4 （RPS6KA4） and high mobility group protein B1 （HMGB1）. These findings suggest that tryptanthrin can inhibit breast cancer in mice mainly through regulating tumor inflammatory microenvironment.

关键词

Keywords

references

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