中国新药杂志

西达本胺对小鼠三阴性乳腺癌4T-1细胞增殖及上皮间质转化的影响

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任雪 ¹ , 陈倩影 ¹ , 张浩 ¹ , 李银芳 ¹ , 杨盛力 ² , 杜志勇 ³ , 曹春雨 ¹

中国新药杂志 | 实验研究 2021,30(23): 2215-2224

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中国新药杂志 |实验研究 2021 , 30 (23) : 2215 -2224

西达本胺对小鼠三阴性乳腺癌4T-1细胞增殖及上皮间质转化的影响

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任雪¹, 陈倩影¹, 张浩¹, 李银芳¹, 杨盛力², 杜志勇³, 曹春雨¹

作者信息

¹三峡大学医学院，肿瘤微环境与免疫治疗湖北省重点实验室，宜昌　443002

²华中科技大学同济医学院附属协和医院，武汉　430022

³三峡大学人民医院，乳腺甲状腺小儿外科，宜昌　443002

通讯作者:

曹春雨，男，副教授，主要从事肿瘤表观遗传学研究。联系电话：（0717）6397179，E-mail：caocy@ctgu.edu.cn。

作者简介:

任雪，女，硕士研究生，主要从事分子生物学与肿瘤免疫学研究。联系电话：（0717）6397179，E-mail：1250129932@qq.com。

Effect of chidamide on proliferation and epithelial mesenchymal transformation of murine triple negative breast cancer 4T-1 cells

Xue REN¹, Qian-ying CHEN¹, Hao ZHANG¹, Yin-fang LI¹, Sheng-li YANG², Zhi-yong DU³, Chun-yu CAO¹

Affiliations

¹Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, China Three Gorges University, Yichang 443002, China

²Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

³Renmin Hospital of China Three Gorges University, Breast Thyroid Pediatric Surgery, Yichang 443002, China

出版时间: 2021-12-15

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摘要

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目的：

探索组蛋白去乙酰化酶抑制剂西达本胺（chidamide）对三阴性小鼠乳腺癌4T-1细胞增殖、侵袭、迁移及上皮间质转化（epithelial-mesenchymal transition，EMT）的影响。

方法：

CCK-8法分析西达本胺对4T-1细胞增殖的影响；流式细胞术检测西达本胺对4T-1细胞的细胞周期和细胞凋亡的影响；Western Blot检测细胞内组蛋白去乙酰化酶5（histone deacetylase 5，HDAC5）、赖氨酸特异性去甲基化酶1（lysine-specifc demethylase 1，LSD1）以及EMT相关蛋白E-cadherin、波形蛋白（vimentin）、MMP-9和MMP-2的表达；Transwell法结合结晶紫染色分析西达本胺对4T-1细胞迁移、侵袭的影响；HE染色法观察肿瘤组织的肺部转移情况及西达本胺对小鼠的毒性作用；ELISA法分析西达本胺对小鼠免疫系统的影响。

结果：

CCK-8法检测结果显示西达本胺具有显著抑制4T-1细胞增殖的能力，其24 h的IC₅₀值为43.42 μmol·L^-1。西达本胺可诱导4T-1细胞发生G₂/M期阻滞以及细胞凋亡。同时发现，西达本胺显著下调4T-1细胞中HDAC5的mRNA和蛋白表达，但只下调LSD1的蛋白表达；此外，4T-1细胞内EMT相关因子E-cadherin，vimentin，MMP-9和MMP-2的表达显著下调。Transwell实验发现，西达本胺显著抑制4T-1细胞的迁移和侵袭。体内实验表明，10 mg·kg^-1西达本胺抑制了4T-1细胞在荷瘤小鼠体内的增殖和肺转移，未对小鼠体重及心脏、肝脏、肾脏组织形态产生显著影响。

结论：

西达本胺能够下调HDAC5，LSD1和EMT相关蛋白E-cadherin，vimentin，MMP-9和MMP-2的表达，并抑制4T-1细胞体内、体外的增殖与迁移，提示西达本胺具有用于临床抗三阴性乳腺癌治疗的潜在价值。

关键词

西达本胺 / 组蛋白去乙酰化酶5 / 三阴性乳腺癌 / 上皮间质转化 / 肿瘤转移

Abstract

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Objective:

To explore the effects of histone deacetylase inhibitor chidamide on the proliferation, invasion, migration and epithelial-mesenchymal transition (EMT) of murine triple-negative breast cancer 4T-1 cells.

Methods:

CCK-8 method was performed to analyze the effects of chidamide on the proliferation of 4T-1 cells; flow cytometry to detect the effects of chidamide on the cell cycle and apoptosis of 4T-1 cells; Western Blot to detect intracellular histone deacetylase 5, lysine-specifc demethylase 1 and EMT-related proteins E-cadherin, Vimentin, MMP-9, MMP-2 expression; Transwell method to analyze the effect of chidamide on the migration and invasion of 4T-1 cells; HE staining method to observe the pulmonary metastasis of tumor tissues and the toxicity of chidamide on mice; ELISA method to analyze the effect of chidamide on the immune system in mice.

Results:

The results of CCK-8 method showed that chidaniline had the ability to prominently inhibit the proliferation of 4T-1 cells, and its 24 h IC₅₀ value was 43.42 μmol·L^-1. Chidamide could induce G₂/M phase arrest and apoptosis of 4T-1 cells. At the same time, it was found that chidamide distinctly down-regulated the mRNA and protein expression of HDAC5 in 4T-1 cells, but only down-regulated the protein expression of LSD1; in addition, the expressions of EMT-related factors E-cadherin, vimentin, MMP-9, MMP-2 were notably down-regulated. Transwell experiments found that chidamide obviously inhibited the migration and invasion of 4T-1 cells. In vivo, chidamide of 10 mg·kg^-1 showed an outstanding suppression effect on the proliferation and pulmonary metastasis of 4T-1 cells in tumor-bearing mice, but did not affect the body weight and the morphology of heart, liver, and kidney tissues.

Conclusion:

Chidamide can depress the expression of HDAC5, LSD1 and EMT-related proteins E-cadherin, Vimentin, MMP-9, MMP-2, and inhibit the proliferation and migration of 4T-1 cells both in vivo and in vitro, indicating that chidamide has a great potential for clinical anti-triple-negative breast cancer treatment.

Key words

chidamide / histone deacetylase 5 / triple-negative breast cancer / epithelial-mesenchymal transition / tumor metastasis

引用本文

任雪, 陈倩影, 张浩, 李银芳, 杨盛力, 杜志勇, 曹春雨. 西达本胺对小鼠三阴性乳腺癌4T-1细胞增殖及上皮间质转化的影响. 中国新药杂志, 2021 , 30 (23) : 2215 -2224 .

Xue REN, Qian-ying CHEN, Hao ZHANG, Yin-fang LI, Sheng-li YANG, Zhi-yong DU, Chun-yu CAO. Effect of chidamide on proliferation and epithelial mesenchymal transformation of murine triple negative breast cancer 4T-1 cells[J]. Chinese Journal of New Drugs, 2021 , 30 (23) : 2215 -2224 .

基金

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国家自然科学基金面上项目(81772833)

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2021年第30卷第23期

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首发时间：2026-10-28
出版时间：2021-12-15

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录用日期：2020-09-17

基金

国家自然科学基金面上项目(81772833)

作者信息

¹三峡大学医学院，肿瘤微环境与免疫治疗湖北省重点实验室，宜昌　443002

²华中科技大学同济医学院附属协和医院，武汉　430022

³三峡大学人民医院，乳腺甲状腺小儿外科，宜昌　443002

通讯作者:

曹春雨，男，副教授，主要从事肿瘤表观遗传学研究。联系电话：（0717）6397179，E-mail：caocy@ctgu.edu.cn。

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https://castjournals.cast.org.cn/joweb/zgxyzz/CN/1322127355712225438

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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