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miR-362-3p 通过靶向调控垂体瘤转化基因 1 抑制口腔鳞状细胞癌细胞的侵袭和转移。

miR-362-3p inhibited the invasion and metastasis of oral squamous cell carcinoma cells by targeting the regulation of pituitary tumor-transforming gene 1.

机构信息

Dept. of Stomatology, Affiliated Hospital of Weifang Medical University, Weifang 261000, China.

School of Stomatology, Weifang Medical University, Weifang 261053, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2024 Feb 1;42(1):46-55. doi: 10.7518/hxkq.2024.2023237.

Abstract

OBJECTIVES

This study aimed to explore the effect of pituitary tumor-transforming gene 1 (PTT-G1) on the invasion and proliferation of oral squamous cell carcinoma (OSCC) cell lines under the action of miR-362-3p.

METHODS

The bioinformatics online database was used to query the expression of PTTG1 in head and neck squamous cell carcinoma (HNSCC). The expression of PTTG1 in the Cal-27, HN-30, and HOK cell lines was detected by Western blot. A wound-healing assay was used to determine the effect of PTTG1 on the migration ability of the OSCC cells. The Transwell assay was used to examine the changes in cell-invasion ability. 5-ethynyl-2'-deoxyuridine (EdU) cell-proliferation assay was used to detect changes in cell-proliferation ability. Bioinformatics approach predicted the upstream miRNA of PTTG1. The targeting relationship between miR-362-3p and PTTG1 was examined by the dual luciferase assay, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of miRNA in OSCC tissues.

RESULTS

The ENCORI database showed that PTTG1 expression was up-regulated in OSCC tissues. Western blot confirmed that PTTG1 expression was up-regulated in Cal-27 and HN-30 cells than HOK cells. PTTG1 knockout can inhibit the migration, invasion, and proliferation of Cal-27 and HN-30 cells (<0.05). Bioinformatics prediction websites predicted that the upstream miRNA of PTTG1 was miR-362-3p, and PTTG1 can bind to miR-362-3p. Results of qRT-PCR showed that miR-362-3p expression was downregulated in OSCC tissues compared with normal tissue (<0.05). Transwell and EdU experiments confirmed that miR-362-3p knockdown can promote the invasion and proliferation of Cal-27 and HN-30 after PTTG1 knockdown.

CONCLUSIONS

miR-362-3p can inhibit the invasion and proliferation of Cal-27 and HN-30 cells by targeting PTTG1.

摘要

目的

本研究旨在探讨垂体肿瘤转化基因 1(PTTG1)在 miR-362-3p 作用下对口腔鳞状细胞癌细胞系侵袭和增殖的影响。

方法

利用生物信息学在线数据库查询头颈部鳞状细胞癌(HNSCC)中 PTTG1 的表达情况。采用 Western blot 检测 Cal-27、HN-30 和 HOK 细胞系中 PTTG1 的表达。通过划痕实验测定 PTTG1 对 OSCC 细胞迁移能力的影响。Transwell 实验检测细胞侵袭能力的变化。5-乙炔基-2'-脱氧尿苷(EdU)细胞增殖实验检测细胞增殖能力的变化。生物信息学方法预测 PTTG1 的上游 miRNA。通过双荧光素酶报告基因实验检测 miR-362-3p 与 PTTG1 的靶向关系,并用定量实时聚合酶链反应(qRT-PCR)检测 OSCC 组织中 miRNA 的表达。

结果

ENCORI 数据库显示 PTTG1 在 OSCC 组织中表达上调。Western blot 证实 PTTG1 在 Cal-27 和 HN-30 细胞中的表达高于 HOK 细胞。PTTG1 敲除可抑制 Cal-27 和 HN-30 细胞的迁移、侵袭和增殖(<0.05)。生物信息学预测网站预测 PTTG1 的上游 miRNA 是 miR-362-3p,PTTG1 可以与 miR-362-3p 结合。qRT-PCR 结果显示,与正常组织相比,OSCC 组织中 miR-362-3p 的表达下调(<0.05)。Transwell 和 EdU 实验证实,PTTG1 敲除后 miR-362-3p 表达下调可促进 Cal-27 和 HN-30 侵袭和增殖。

结论

miR-362-3p 可通过靶向 PTTG1 抑制 Cal-27 和 HN-30 细胞的侵袭和增殖。

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