miR-362-3p调控垂体肿瘤转化基因1抑制口腔鳞状细胞癌侵袭及增殖的研究

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

摘要

目的 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（OSCC）细胞Cal-27、HN-30侵袭以及增殖能力的影响。 方法 生物信息学在线数据库查询PTTG1在头颈部鳞状细胞癌（HNSCC）中的表达。蛋白质印迹法（Western blot）实验检测PTTG1在Cal-27、HN-30以及HOK细胞系中的表达。划痕愈合实验、Transwell侵袭实验及5-乙炔基-2’脱氧尿嘧啶核苷（EdU）细胞增殖实验检测PTTG1对Cal-27、HN-30细胞迁移、侵袭、增殖的影响。生物信息学在线数据库预测PTTG1的上游miRNA，双荧光素酶实验检测结合情况，实时荧光定量聚合酶链反应（qRT-PCR）检测该miRNA在组织中的表达。 结果 ENCORI数据库结果显示PTTG1在OSCC组织中表达上调；Western blot实验显示Cal-27、HN-30细胞中PTTG1表达量较HOK细胞中高。转染Si-PTTG1质粒的Cal-27、HN-30细胞的迁移能力、侵袭能力和细胞增殖能力均较对照组降低（P<0.05）。通过网站预测出PTTG1的上游miRNA为miR-362-3p，双荧光素酶实验检测出PTTG1与miR-362-3p存在结合位点；qRT-PCR检测结果显示miR-362-3p在OSCC肿瘤组织中相对于正常组织表达下调（P<0.05）；并且敲低miR-362-3p的表达后能够促进敲低PTTG1后的Cal-27、HN-30侵袭和增殖。 结论 miR-362-3p可通过靶向PTTG1抑制Cal-27、HN-30细胞侵袭、增殖。

Abstract

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

关键词

口腔鳞状细胞癌 / 垂体肿瘤转化基因1 / 微小RNA / 侵袭 / 增殖

Key words

oral squamous cell carcinoma / pituitary tumor-transforming gene 1 / microRNA / invasion / proliferation

Author summay

丁啸，住院医师，硕士，E-mail：[e-mail]