Department of Otorhinolaryngology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
J Physiol Pharmacol. 2022 Apr;73(2). doi: 10.26402/jpp.2022.2.03. Epub 2022 Aug 18.
Laryngeal squamous cell carcinoma (LSCC) is diagnosed as a malignant tumor with a poor prognosis, the associated mechanisms still need to be further investigated. The LINC01554 gene is confirmed to participate in the tumorigenesis of hepatocellular carcinoma, but its role in LSCC has not been investigated. The aim of this study was to investigate the function and the potential mechanism of LINC01554 in LSCC, LINC01554 further was used as a molecular target for the diagnosis and molecular targeted therapy of LSCC. The microarray-based gene expression profiling of LSCC and its adjacent non-tumor tissue were used to identify the differentially expressed long non-coding RNAs (lncRNAs). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to verify the expression levels of LINC01554 in tissue and LSCC cell lines. The DNA methylation level of the LINC01554 promoter was detected by the application of bisulfite genomic sequencing (BGS), and the bisulfite conversion-specific/methylation-specific polymerase chain reaction (BS-MSP) method. The effect of LINC01554 on the proliferation, migration, and invasion of squamous cell carcinoma was assessed by MTS, wound healing, transwell, RT-qPCR, Western blot in vitro-cultured TU177 cells, and AMC-HN-8 cells. The microarray-based gene expression profiling identified the differentially expressed lncRNAs, including LINC01554, with downregulation in the LSCC tissue vs. normal tissue. The RT-qPCR verified the downregulation of LINC01554 in the LSCC tissue (P=0.0049) and LSCC cell (P=0.0020). The BGS and BS-MSP exhibited the hypermethylation level of the LINC01554 promoter, which mediated the downregulation of LINC01554. A gain-of-function experiment showed that LINC01554 inhibited the proliferation, migration, and invasion of TU177 and AMC-HN-8. Subsequently, LINC01554 overexpression was shown to decrease cell viability in TU177 and AMC-HN-8 cells treated with cisplatin. Our findings indicated that the aberrant methylation-mediated downregulation of LINC01554 promoted malignant progression and cisplatin resistance in LSCC, and LINC01554 may serve as a potential diagnostic biomarker and a novel therapeutic target for LSCC.
喉鳞状细胞癌（LSCC）被诊断为预后不良的恶性肿瘤，其相关机制仍需进一步研究。LINC01554 基因已被证实参与肝癌的肿瘤发生，但它在 LSCC 中的作用尚未被研究。本研究旨在探讨 LINC01554 在 LSCC 中的功能和潜在机制，并将其进一步用作 LSCC 的诊断和分子靶向治疗的分子靶点。
基于微阵列的 LSCC 及其相邻非肿瘤组织的基因表达谱用于鉴定差异表达的长非编码 RNA（lncRNA）。逆转录定量聚合酶链反应（RT-qPCR）用于验证组织和 LSCC 细胞系中 LINC01554 的表达水平。应用亚硫酸氢盐基因组测序（BGS）检测 LINC01554 启动子的 DNA 甲基化水平，并应用亚硫酸氢盐转化特异性/甲基化特异性聚合酶链反应（BS-MSP）方法。
在体外培养的 TU177 细胞和 AMC-HN-8 细胞中，通过 MTS、划痕愈合、Transwell、RT-qPCR、Western blot 评估 LINC01554 对鳞状细胞癌增殖、迁移和侵袭的影响。基于微阵列的基因表达谱鉴定了差异表达的 lncRNA，包括 LINC01554，在 LSCC 组织与正常组织相比下调。RT-qPCR 验证了 LSCC 组织（P=0.0049）和 LSCC 细胞（P=0.0020）中 LINC01554 的下调。BGS 和 BS-MSP 显示 LINC01554 启动子的高甲基化水平，介导 LINC01554 的下调。功能获得实验表明，LINC01554 抑制 TU177 和 AMC-HN-8 的增殖、迁移和侵袭。随后，在 TU177 和 AMC-HN-8 细胞中用顺铂处理时，LINC01554 的过表达显示降低细胞活力。
我们的研究结果表明，异常甲基化介导的 LINC01554 下调促进了 LSCC 的恶性进展和顺铂耐药，LINC01554 可能作为 LSCC 的潜在诊断生物标志物和新型治疗靶点。
