Jin Long, Pan Yu-Liang, Zhang Jun, Cao Pei-Guo
Department of Oncology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China.
Department of Oncology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China.
Transl Oncol. 2021 Apr;14(4):101036. doi: 10.1016/j.tranon.2021.101036. Epub 2021 Feb 12.
Colorectal cancer causes severe burdensome on the health by its high fatality and poor prognosis. Hox transcript antisense intergenic RNA (HOTAIR) was believed closely related with the genesis and development of colorectal cancer, but the regulatory mechanism is still to be investigated. The expression of HOTAIR was analyzed in colorectal cancer using both qRT-PCR and ISH assay. The cell viability, migration, invasion and apoptosis rate were evaluated using MTT, BrdU,Transwell and flow cytometryexperiments. The interaction between HOTAIR and SNAIL was detected using RIP and RNA pull-down. The binding of SNAIL to HNF4α promoter was assessed by ChIP. The cell lines that knock down HOTAIR, SNAIL or overexpress HNF4α were constructed using retroviral vector system. The tumorigenic and metastatic capacity of colorectal cancer cells after knocking down HOTAIR were evaluated based on xenograft assay and liver metastases model. HOTAIR was highly expressed in both tissue and cell lines of colorectal cancer, indicated a regulatory function in colorectal cancer. Knock-down of HOTAIR suppressed cell viability, migration, invasion and epithelial-mesenchymal transition (EMT) of colorectal cancer cells in vitro, and inhibited the growth and metastasis of colorectal tumor in nude mice. We further found that HOTAIR suppressed HNF4α via recruiting SNAIL, and the overexpression of HNF4α inhibited cell viability, migration, invasion and EMT of colorectal cancer cells. We demonstrated that HOTAIR regulates the level of HNF4α via recruiting SNAIL, knocking down HOTAIR repressed the cell viability and metestasis of colorectal cancer cell line in vitro, and suppressed the tomorgenesis and migration/invasion of colorectal cancer in vivo.
结直肠癌因其高死亡率和不良预后给健康带来了严重负担。Hox转录反义基因间RNA（HOTAIR）被认为与结直肠癌的发生和发展密切相关，但其调控机制仍有待研究。使用qRT-PCR和ISH检测分析了HOTAIR在结直肠癌中的表达。使用MTT、BrdU、Transwell和流式细胞术实验评估细胞活力、迁移、侵袭和凋亡率。使用RIP和RNA下拉检测HOTAIR与SNAIL之间的相互作用。通过ChIP评估SNAIL与HNF4α启动子的结合。使用逆转录病毒载体系统构建敲低HOTAIR、SNAIL或过表达HNF4α的细胞系。基于异种移植实验和肝转移模型评估敲低HOTAIR后结直肠癌细胞的致瘤和转移能力。HOTAIR在结直肠癌的组织和细胞系中均高表达，表明其在结直肠癌中具有调控功能。敲低HOTAIR可在体外抑制结直肠癌细胞的活力、迁移、侵袭和上皮-间质转化（EMT），并在裸鼠中抑制结直肠肿瘤的生长和转移。我们进一步发现，HOTAIR通过招募SNAIL抑制HNF4α，而HNF4α的过表达可抑制结直肠癌细胞的活力、迁移、侵袭和EMT。我们证明，HOTAIR通过招募SNAIL调节HNF4α的水平，敲低HOTAIR可在体外抑制结直肠癌细胞系的活力和转移，并在体内抑制结直肠癌的发生和迁移/侵袭。
