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

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

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。

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.