口腔医学研究

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (7): 640-647.DOI: 10.13701/j.cnki.kqyxyj.2024.07.013

• 口腔黏膜病学研究 • 上一篇    下一篇

miR-24通过调控AKT/β-catenin信号通路及EMT过程对涎腺腺样囊性癌细胞生物学功能的影响及机制

李琳1, 姬晓霖1, 姜向瑞2*   

  1. 1.山西白求恩医院(山西医学科学院),山西医科大学第三医院;同济山西医院耳鼻咽喉头颈外科 山西 太原 030032;
    2.山西齿科医院种植科 山西 太原 030027
  • 收稿日期:2024-03-26 出版日期:2024-07-28 发布日期:2024-07-24
  • 通讯作者: *姜向瑞,E-mail:yayijxr@163.com
  • 作者简介:李琳(1985~ ),女,山西运城人,硕士,主治医师,主要从事侧颅底肿瘤相关研究。
  • 基金资助:
    山西省基础研究计划青年科学研究项目(编号:202203021212107)

Effects and Mechanisms of miR-24 on Biological Functions of Salivary-gland Adenoid Cystic Cancer Cells through Regulation of AKT/β-catenin Signaling Pathway and EMT Process

LI Lin1, JI Xiaolin1, JIANG Xiangrui2*   

  1. 1. Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Department of Otolaryngology Head and Neck Surgery, Tongji Shanxi Hospital, Taiyuan 030032, China;
    2. Department of Dental Implant, Shanxi Dental Hospital, Taiyuan 030027, China
  • Received:2024-03-26 Online:2024-07-28 Published:2024-07-24

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摘要： 目的:探究微小RNA(microRNA,miR)-24通过调控蛋白激酶B(protein kinase B,AKT)/β-连环蛋白(β-catenin)信号通路及上皮间充质转化(epithhelial-mesenchymal transition,EMT)过程对涎腺腺样囊性癌细胞生物学功能的影响及机制。方法:通过收集2021年6月~2024年3月本院收治的60例涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC)患者。苏木精-伊红(hematoxylin-eosin,HE)染色检测SACC组织病理类型。培养人SACC细胞系(SACC-83和SACC-LM),通过荧光原位杂交(fluorescence in situ hybridization,FISH)及实时荧光定量逆转录聚合酶链反应(quantitative real time polymerase chain reaction,qRT-PCR)检测SACC组织及细胞中miR-24表达。通过细胞转染将miR-24抑制剂(miR-24 inhibitor)转染至SACC细胞中,通过细胞计数试剂盒-8(cell count kit-8,CCK-8)、克隆形成实验、细胞划痕实验、Transwell实验检测SACC细胞生物学行为,Western blot检测上皮间充质转化(EMT)相关蛋白及AKT/β-catenin信号通路蛋白表达情况。结果:HE染色结果发现,本研究SACC患者癌组织包含3种主要病理类型:实体型、筛状型和管状型。与正常组织相比,miR-24在SACC组织、SACC-83细胞和SACC-LM细胞中高表达,且miR-24在SACC-LM中的表达程度高于SACC-83。miR-24 inhibitor显著抑制SACC-83和SACC-LM的细胞活力、细胞克隆数量、细胞划痕细胞迁移率、迁移、侵袭。miR-24 inhibiotr转染后SACC-83和SACC-LM细胞中上皮细胞钙粘蛋白(epithelial-cadherin,E-cadherin)水平明显升高,而波形蛋白(Vimentin)和神经钙粘蛋白(neural-cadherin,N-cadherin)水平降低。结论:miR-24通过调控AKT/β-catenin信号通路调节SACC细胞增殖、迁移、侵袭及EMT过程。

关键词: 涎腺腺样囊性癌, 微小RNA-24, 蛋白激酶B/β-连环蛋白, 上皮间充质转化, 迁移, 增殖

Abstract: Objective: To explore the effect and mechanism of microRNA (miR)-24 on the biological function of salivarium adenoid cystic cancer (SACC) cells by regulating protein kinase B (AKT)/β-catenin signaling pathway and EMT process. Methods: Sixty patients with SACC admitted to our hospital from June 2021 to March 2024 were collected, and human SACC cell lines (SACC-83 and SACC-LM) were cultured. The histopathological types of SACC were detected by HE staining. The expression of miR-24 in SACC tissues and cells was detected by fluorescence in situ hybridization (FISH) and quantitative real time polymerase chain reaction (qRT-PCR). miR-24 inhibitor was transfected into SACC cells by cell transfection, and the biological behavior of SACC cells was detected by CCK-8, clonal formation assay, wound healing assay, and transwell assay. Western blot analysis was performed to detect the expression of EMT-related proteins and AKT/β-catenin signaling pathway. Results: HE staining results showed that the cancer tissue of SACC patients contained three main pathological types: solid type, cribriform type, and tubular type. Compared with paracancer tissues, miR-24 was highly expressed in SACC tissues, SACC-83 cells, and SACC-LM cells, and the expression level of miR-24 in SACC-LM was higher than that in SACC-83. miR-24 inhibitor significantly inhibited cell viability, colony formation, wound healing rate, migration, and invasion of SACC-83 and SACC-LM. After transfection of miR-24 inhibitor, E-cadherin levels were significantly increased in SACC-83 and SACC-LM cells, while Vimentin and N-cadherin levels were decreased. Conclusion: miR-24 regulates SACC cell proliferation, migration, invasion, and EMT process by regulating AKT/β-catenin signaling pathway.

Key words: salivary adenoid cystic carcinoma, miR-24, AKT/β-catenin, epithelial mesenchymal transformation, migration, proliferation