Song Fucun, Yang Yang, Liu Jixiang
Department of Otolaryngology Head and Neck Surgery, Tianjin Union Medical Center, Tianjin 300121, P.R. China.
Exp Ther Med. 2021 Mar;21(3):232. doi: 10.3892/etm.2021.9663. Epub 2021 Jan 21.
Accumulating evidence indicates that the long non-coding RNA myocardial infarction associated transcript (lncRNA MIAT) serves an important role in the progression of a number of cancer types. However, the precise molecular mechanism of MIAT in laryngeal squamous cell carcinoma (LSCC) progression remain elusive. The aim of the current study was to assess the effects and to clarify the molecular mechanism of MIAT on the proliferation and invasion of LSCC cells. The expression of MIAT was detected in LSCC tissues and cells using reverse transcription-quantitative PCR. MTT and colony formation assays were performed to examine the effects of MIAT on the proliferation of LSCC cells. Additionally, wound healing and Transwell experiments were employed to examine cellular migration and invasion. Luciferase reporter gene assay was also used to confirm the direct binding between MIAT and microRNA (miR)-613 in LSCC cells. An RNA immunoprecipitation assay was performed to verify the interaction between MIAT and miR-613. In the present study, it was found that the expression of MIAT in LSCC tissues was markedly higher compared with that in adjacent non-tumor tissues. In addition, MIAT expression was also increased in the human LSCC cell lines TU686, TU-177 and AMC-HN-8 compared with that in normal human keratinocytes (HaCaT). Knocking down MIAT expression significantly reduced LSCC cell proliferation and inhibited colony formation, a shown by MTT and colony formation assays, respectively. MIAT knockdown also substantially inhibited the migratory and invasive abilities of LSCC cells, as shown by wound healing and Transwell invasion assays, respectively. Subsequently, luciferase reporter assays verified that MIAT could bind to miR-613, where a negative correlation was observed between the expression of MIAT and miR-613 in LSCC tissues. Suppression of miR-613 partially reversed the inhibitory effects of MIAT knockdown on the proliferation, migration and invasion of LSCC cells. Taken together, the present study identified that MIAT may function as an oncogenic lncRNA to promote LSCC progression, which provides a potential therapeutic target or as a novel diagnostic biomarker for LSCC.
越来越多的证据表明,长链非编码RNA心肌梗死相关转录本(lncRNA MIAT)在多种癌症类型的进展中发挥重要作用。然而,MIAT在喉鳞状细胞癌(LSCC)进展中的精确分子机制仍不清楚。本研究的目的是评估MIAT对LSCC细胞增殖和侵袭的影响,并阐明其分子机制。采用逆转录定量PCR检测LSCC组织和细胞中MIAT的表达。进行MTT和集落形成试验以检测MIAT对LSCC细胞增殖的影响。此外,采用伤口愈合试验和Transwell试验检测细胞迁移和侵袭能力。还使用荧光素酶报告基因试验来证实LSCC细胞中MIAT与微小RNA(miR)-613之间的直接结合。进行RNA免疫沉淀试验以验证MIAT与miR-613之间的相互作用。在本研究中,发现LSCC组织中MIAT的表达明显高于相邻的非肿瘤组织。此外,与正常人角质形成细胞(HaCaT)相比,人LSCC细胞系TU686、TU-177和AMC-HN-8中MIAT的表达也增加。MTT和集落形成试验分别显示,敲低MIAT表达显著降低LSCC细胞增殖并抑制集落形成。伤口愈合试验和Transwell侵袭试验分别显示,敲低MIAT也显著抑制LSCC细胞的迁移和侵袭能力。随后,荧光素酶报告试验证实MIAT可以与miR-613结合,并且在LSCC组织中MIAT与miR-613的表达之间存在负相关。抑制miR-613可部分逆转敲低MIAT对LSCC细胞增殖、迁移和侵袭的抑制作用。综上所述,本研究确定MIAT可能作为一种致癌lncRNA促进LSCC进展,这为LSCC提供了一个潜在的治疗靶点或作为一种新的诊断生物标志物。