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circ_0005379 通过调控 miR-17-5p/酰基辅酶 A 氧化酶 1 轴抑制口腔鳞状细胞癌的进展。

circ_0005379 inhibits the progression of oral squamous cell carcinoma by regulating the miR-17-5p/acyl-CoA oxidase 1 axis.

机构信息

Center of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Aug 1;39(4):425-433. doi: 10.7518/hxkq.2021.04.008.

Abstract

OBJECTIVES

To investigate the effects of circ_0005379 on the proliferation, apoptosis, migration, and invasion of oral squamous cell carcinoma (OSCC) cells and its mechanism.

METHODS

Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression levels of circ_0005379 and miR-17-5p in OSCC tissues and SCC15 cell lines. Western blot was used to detect the expression levels of acyl-CoA oxidase 1 (ACOX1). The circ_0005379 overexpression vector was transfected into SCC15 cells. Methyl thiazolyl tetrazolium blue staining, flow cytometry, Transwell, and Western blot were used to detect the effects of circ_0005379 overexpression on the proliferation, apoptosis, migration, and invasion of SCC15 cells and the expression of E-cadherin, β-catenin, and Snail proteins. Dual luciferase reporter assay and RNA immunoprecipitation were used to examine the regulation of circ_0005379, miR-17-5p, miR-17-5p, and ACOX1 in SCC15 cells. A nude mouse xenograft model of SCC15 cells stably overexpressing circ_0005379 was established, and the effect of circ_0005379 overexpression on the growth of xenografts in nude mice was observed.

RESULTS

Compared with adjacent cancer tissues, the expression levels of circ_0005379 and ACOX1 proteins in OSCC tissues were decreased (<0.05), and the expression level of miR-17-5p was increased (<0.05). Compared with HOK-16A cells, the expression levels of circ_0005379 and ACOX1 proteins in SCC15 cell lines were decreased (<0.05), and the expression level of miR-17-5p was increased (<0.05). After overexpressing circ_0005379, the activity and number of migrating and invading SCC15 cells and the expression levels of β-catenin and Snail proteins were decreased (<0.05); however, the apoptosis rate and expression level of E-cadherin protein were increased (<0.05). In SCC15 cells, circ_0005379 targeted the negative regulation of miR-17-5p expression, and miR-17-5p targeted the negative regulation of ACOX1 expression. Overexpressing miR-17-5p or silencing ACOX1 could reverse the effects of circ_0005379 overexpression on the proliferation, apoptosis, migration, and invasion of OSCC cell lines. The tumor volume and weight of nude mice overexpressing circ_0005379 were decreased (<0.05), the expression levels of circ_0005379 and ACOX1 protein in tumor tissues were increased (<0.05), and the expression level of miR-17-5p was decreased (<0.05).

CONCLUSIONS

circ_0005379 may inhibit the proliferation, migration, and invasion of OSCC cells by downregulating the expression of miR-17-5p and upregulating ACOX1, which promote apoptosis and inhibit tumor growth . circ_0005379 may be a potential target for OSCC treatment.

摘要

目的

探讨 circ_0005379 对口腔鳞状细胞癌(OSCC)细胞增殖、凋亡、迁移和侵袭的影响及其机制。

方法

实时定量聚合酶链反应(RT-qPCR)检测 OSCC 组织和 SCC15 细胞系中 circ_0005379 和 miR-17-5p 的表达水平。Western blot 检测酰基辅酶 A 氧化酶 1(ACOX1)的表达水平。将 circ_0005379 过表达载体转染至 SCC15 细胞。噻唑蓝比色法、流式细胞术、Transwell 实验和 Western blot 检测 circ_0005379 过表达对 SCC15 细胞增殖、凋亡、迁移和侵袭的影响以及 E-钙黏蛋白、β-连环蛋白和 Snail 蛋白的表达。双荧光素酶报告基因实验和 RNA 免疫沉淀检测 SCC15 细胞中 circ_0005379、miR-17-5p、miR-17-5p 和 ACOX1 的调控作用。建立 SCC15 细胞稳定过表达 circ_0005379 的裸鼠异种移植模型,观察 circ_0005379 过表达对裸鼠异种移植瘤生长的影响。

结果

与癌旁组织相比,OSCC 组织中 circ_0005379 和 ACOX1 蛋白的表达水平降低(<0.05),miR-17-5p 的表达水平升高(<0.05)。与 HOK-16A 细胞相比,SCC15 细胞系中 circ_0005379 和 ACOX1 蛋白的表达水平降低(<0.05),miR-17-5p 的表达水平升高(<0.05)。过表达 circ_0005379 后,SCC15 细胞的活性和迁移、侵袭细胞数以及β-连环蛋白和 Snail 蛋白的表达水平降低(<0.05);而 E-钙黏蛋白蛋白的表达水平升高(<0.05),细胞凋亡率升高。在 SCC15 细胞中,circ_0005379 靶向负调控 miR-17-5p 的表达,miR-17-5p 靶向负调控 ACOX1 的表达。过表达 miR-17-5p 或沉默 ACOX1 可逆转 circ_0005379 过表达对 OSCC 细胞系增殖、凋亡、迁移和侵袭的影响。过表达 circ_0005379 的裸鼠肿瘤体积和重量减小(<0.05),肿瘤组织中 circ_0005379 和 ACOX1 蛋白的表达水平升高(<0.05),miR-17-5p 的表达水平降低(<0.05)。

结论

circ_0005379 可能通过下调 miR-17-5p 的表达和上调 ACOX1 来抑制 OSCC 细胞的增殖、迁移和侵袭,促进细胞凋亡并抑制肿瘤生长。circ_0005379 可能是 OSCC 治疗的潜在靶点。

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