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circ0005027 通过调控 miR-548c-3p/CDH1 轴作为 ceRNA 影响下咽鳞癌细胞的恶性生物学行为。

circ0005027 Acting as a ceRNA Affects the Malignant Biological Behavior of Hypopharyngeal Squamous Cell Carcinoma by Modulating miR-548c-3p/CDH1 Axis.

机构信息

The Second Department of Otolaryngology Head and Neck Surgery, First Affiliated Hospital of Kunming Medical University, No. 295 Xichang Rd, Kunming, 650032, Yunnan, China.

Department of Otolaryngology, The First People's Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming, 650034, Yunnan, China.

出版信息

Biochem Genet. 2024 Aug;62(4):2853-2868. doi: 10.1007/s10528-023-10570-y. Epub 2023 Nov 29.

Abstract

Hypopharyngeal squamous cell carcinoma (HSCC) is a malignant tumor of head and neck. It was verified that circ0005027 was downregulated in HSCC tissues. Here, we aimed to investigate the function and specific regulatory mechanism of circ0005027 in HSCC. Ten pairs tissues of HSCC and adjacent para-cancer were collected. Reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) measured circ0005027, miR-548c-3p, and Cadherin 1 (CDH1) mRNA expression. CCK-8 analyzed cell proliferation viability. Flow cytometry assay detected cell cycle and apoptosis rate. Clonal formation assay measured the clonal ability. Transwell detected cell invasion ability. Western blot was performed to detect CDH1, LAST1, p-LAST1, MST1, p-MST1, YAP1, p-YAP1, TAZ and p-TAZ protein level. Dual-luciferase, RIP and RNA pull-down assay identified the target relationship among circ0005027, miR-548c-3p and CDH1. circ0005027 was decreased in tissues and FaDu cells of HSCC. Overexpression of circ0005027 inhibited cell viability, G1-S transition, clonal formation, and invasion and increased cell apoptosis. circ0005027 acted as a ceRNA and decreased circ0005027 enhanced the malignant process of FaDu cells through sponging miR-548c-3p and inhibiting CDH1 expression. Overexpression of CDH1 activated YAP1/TAZ pathway and inhibited the growth of HSCC in vitro. circ0005027 might act as a potential biomarker for the progression and prognosis prediction in HSCC by regulating miR-548c-3p/CDH1/ YAP1/TAZ signaling pathway.

摘要

下咽鳞状细胞癌 (HSCC) 是一种头颈部恶性肿瘤。已验证 circ0005027 在 HSCC 组织中下调。在这里,我们旨在研究 circ0005027 在 HSCC 中的功能和特定调节机制。收集了 10 对 HSCC 组织和相邻癌旁组织。逆转录定量实时聚合酶链反应 (RT-qPCR) 测量 circ0005027、miR-548c-3p 和钙粘蛋白 1 (CDH1) mRNA 表达。CCK-8 分析细胞增殖活力。流式细胞术检测细胞周期和凋亡率。克隆形成实验检测克隆能力。Transwell 检测细胞侵袭能力。Western blot 检测 CDH1、LAST1、p-LAST1、MST1、p-MST1、YAP1、p-YAP1、TAZ 和 p-TAZ 蛋白水平。双荧光素酶报告基因、RIP 和 RNA 下拉实验鉴定了 circ0005027、miR-548c-3p 和 CDH1 之间的靶关系。circ0005027 在 HSCC 组织和 FaDu 细胞中下调。过表达 circ0005027 抑制细胞活力、G1-S 期转换、克隆形成和侵袭,增加细胞凋亡。circ0005027 作为 ceRNA,通过海绵吸附 miR-548c-3p 并抑制 CDH1 表达,增强 FaDu 细胞的恶性进程。CDH1 的过表达激活 YAP1/TAZ 通路,抑制 HSCC 的体外生长。circ0005027 可能通过调节 miR-548c-3p/CDH1/YAP1/TAZ 信号通路,作为 HSCC 进展和预后预测的潜在生物标志物。

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