Department of Otorhinolaryngology Head and Neck Surgery, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine, Zhongshan, Guangdong 528400, P.R. China.
Department of Otorhinolaryngology Head and Neck Surgery, Guangdong Provincial People's Hospital, Guangzhou, Guangdong 510000, P.R. China.
Mol Med Rep. 2022 Feb;25(2). doi: 10.3892/mmr.2021.12562. Epub 2021 Dec 8.
It has been reported that long non‑coding RNA HLA complex group 18 (HCG18) is involved in the progression of cancer, acting as an oncogenic gene. The aim of the present study was to investigate the mechanism underlying the action of HCG18 in laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC). The expression levels of HCG18, microRNA (miR)‑133b and fibroblast growth factor receptor 1 (FGFR1) in LHSCC tissues and transfected LHSCC cells were evaluated by reverse transcription‑quantitative PCR or immunohistochemistry. The viability, migration and invasion of transfected LHSCC cells were detected by Cell Counting Kit‑8, wound healing and Transwell assays, respectively. The targeting relationships of HCG18, miR‑133b and FGFR1 were predicted by bioinformatics analysis and confirmed using a dual‑luciferase reporter assay. Moreover, the expression levels of FGFR1, phosphorylated (p)‑PI3K, PI3K, p‑AKT, AKT, p53, Bax and Bcl‑2 in transfected LHSCC cells were measured by western blotting. It was found that the expression levels of HCG18 and FGFR1 were upregulated, but those of miR‑133b were downregulated in LHSCC tissues. Short hairpin RNA (sh) HCG18 and miR‑133b mimic inhibited LHSCC cell viability, while enhancing miR‑133b expression. HCG18 could competitively bind with miR‑133b. Moreover, the miR‑133b inhibitor promoted cell viability, migration, invasion and the expression levels of Bcl‑2, p‑PI3K and p‑AKT, but inhibited the expression levels of p53 and Bax, which were abrogated by shHCG18. miR‑133b could competitively bind with FGFR1, and the miR‑133b mimic decreased the expression level of FGFR1 in transfected LHSCC cells. shFGFR1 promoted the expression levels of p53 and Bax, while inhibiting viability, migration, invasion and Bcl‑2, p‑PI3K and p‑AKT expression in LHSCC cells. In conclusion, the current results indicated that HCG18 facilitated the progression of LHSCC by upregulating FGFR1 via miR‑133b. The present study evaluated the mechanism with regards to the action of HCG18 in LHSCC, and these experimental results may provide novel evidence for targeted therapy of LHSCC.
据报道，长非编码 RNA HLA 复合物组 18（HCG18）参与癌症的进展，作为一种致癌基因。本研究旨在探讨 HCG18 在喉和下咽鳞状细胞癌（LHSCC）中的作用机制。通过逆转录定量 PCR 或免疫组织化学法检测 LHSCC 组织和转染的 LHSCC 细胞中 HCG18、微小 RNA（miR）-133b 和成纤维细胞生长因子受体 1（FGFR1）的表达水平。通过细胞计数试剂盒-8、划痕愈合和 Transwell 测定分别检测转染的 LHSCC 细胞的活力、迁移和侵袭。通过生物信息学分析预测 HCG18、miR-133b 和 FGFR1 的靶向关系，并通过双荧光素酶报告基因测定进行验证。此外，通过 Western blot 法测定转染的 LHSCC 细胞中 FGFR1、磷酸化（p）-PI3K、PI3K、p-AKT、AKT、p53、Bax 和 Bcl-2 的表达水平。结果发现，LHSCC 组织中 HCG18 和 FGFR1 的表达上调，而 miR-133b 的表达下调。短发夹 RNA（sh）HCG18 和 miR-133b 模拟物抑制 LHSCC 细胞活力，同时增强 miR-133b 的表达。HCG18 可以与 miR-133b 竞争结合。此外，miR-133b 抑制剂促进细胞活力、迁移、侵袭以及 Bcl-2、p-PI3K 和 p-AKT 的表达水平，但抑制 p53 和 Bax 的表达水平，而 shHCG18 则消除了这种抑制作用。miR-133b 可以与 FGFR1 竞争结合，miR-133b 模拟物降低转染的 LHSCC 细胞中 FGFR1 的表达水平。shFGFR1 促进 LHSCC 细胞中 p53 和 Bax 的表达水平，同时抑制细胞活力、迁移、侵袭以及 Bcl-2、p-PI3K 和 p-AKT 的表达。综上所述，本研究结果表明，HCG18 通过 miR-133b 上调 FGFR1 促进 LHSCC 的进展。本研究评估了 HCG18 在 LHSCC 中的作用机制，这些实验结果可能为 LHSCC 的靶向治疗提供新的证据。
