Li Jinxiao, Hu Man, Liu Na, Li Huarong, Yu Zhaomin, Yan Qian, Zhou Minfeng, Wang Yayuan, Song Yanjuan, Pan Guangtao, Liang Fengxia, Chen Rui
Department of Integrated Traditional Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1227 Jiefang Avenue, Wuhan City, 430022, Hubei Province, China.
Rehabilitation Department of traditional Chinese Medicine, Union Red Cross Hospital, Wuhan, 430015, China.
J Exp Clin Cancer Res. 2020 Nov 17;39(1):248. doi: 10.1186/s13046-020-01720-w.
The mechanism of histone deacetylase 3 (HDAC3) in colorectal cancer (CRC) has already been discussed. However, the feedback loop of HDAC3/microRNA (miR)-296-3p and transforming growth factor β-induced factor 1 (TGIF1) in CRC has not been explained clearly. Thus, the mainstay of this study is to delve out the mechanism of this axis in CRC.
To demonstrate that HDAC3 regulates the miR-296-3p/TGIF1/TGFβ axis and is involved in CRC progression, a series of cell biological, molecular and biochemical approaches were conducted from the clinical research level, in vitro experiments and in vivo experiments. These methods included RT-qPCR, Western blot assay, cell transfection, MTT assay, EdU assay, flow cytometry, scratch test, Transwell assay, dual luciferase reporter gene assay, chromatin immunoprecipitation, nude mouse xenograft, H&E staining and TUNEL staining.
Higher HDAC3 and TGIF1 and lower miR-296-3p expression levels were found in CRC tissues. HDAC3 was negatively connected with miR-296-3p while positively correlated with TGIF1, and miR-296-3p was negatively connected with TGIF1. Depleted HDAC3 elevated miR-296-3p expression and reduced TGIF1 expression, decreased TGFβ pathway-related proteins, inhibited CRC proliferation, invasion, and migration in vitro and slowed down tumor growth and induction of apoptosis in vivo, which were reversed by miR-296-3p knockdown. Restored miR-296-3p suppressed TGIF1 and reduced TGFβ pathway-related proteins, inhibited CRC proliferation, invasion, and migration in vitro and slowed down tumor growth and induction of apoptosis in vivo, which were reversed by TGIF1 overexpression.
This study illustrates that down-regulation of HDAC3 or TGIF1 or up-regulation of miR-296-3p discourages CRC cell progression and slows down tumor growth, which guides towards a novel direction of CRC treatment.
组蛋白去乙酰化酶3(HDAC3)在结直肠癌(CRC)中的作用机制已有相关探讨。然而,CRC中HDAC3/微小RNA(miR)-296-3p与转化生长因子β诱导因子1(TGIF1)的反馈回路尚未完全阐明。因此,本研究的主要目的是深入探究该轴在CRC中的作用机制。
为证明HDAC3调控miR-296-3p/TGIF1/TGFβ轴并参与CRC进展,从临床研究水平、体外实验和体内实验等方面开展了一系列细胞生物学、分子和生化实验方法。这些方法包括逆转录定量聚合酶链反应(RT-qPCR)、蛋白质免疫印迹法、细胞转染、MTT法、EdU法、流式细胞术、划痕试验、Transwell试验、双荧光素酶报告基因试验、染色质免疫沉淀、裸鼠异种移植、苏木精-伊红(H&E)染色和末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)染色。
在CRC组织中发现HDAC3和TGIF1表达水平较高,而miR-296-3p表达水平较低。HDAC3与miR-296-3p呈负相关,与TGIF1呈正相关,且miR-296-3p与TGIF1呈负相关。HDAC3缺失可提高miR-296-3p表达并降低TGIF1表达,减少TGFβ通路相关蛋白,在体外抑制CRC增殖、侵袭和迁移,并在体内减缓肿瘤生长和诱导细胞凋亡,而miR-296-3p敲低可逆转这些作用。恢复miR-296-3p可抑制TGIF1并减少TGFβ通路相关蛋白,在体外抑制CRC增殖、侵袭和迁移,并在体内减缓肿瘤生长和诱导细胞凋亡,而TGIF1过表达可逆转这些作用。
本研究表明,下调HDAC3或TGIF1或上调miR-296-3p可抑制CRC细胞进展并减缓肿瘤生长,为CRC治疗提供了新的方向。