BRD4在喉鳞状细胞癌病灶转移过程中的作用及机制研究

目的:探讨溴样结构域蛋白4 (Bromoid Domain Protein 4,BRD4)在喉鳞状细胞癌病灶转移过程中的作用及其机制。方法:收集本院2016年4月-2018年4月收治的87例喉鳞状细胞癌患者手术标本。qRT-PCR、Western blot和免疫组化染色检测患者肿瘤组织(Tumor Tissue)、癌旁组织(Adjacent Tissue)和正常组织(Normal Tissue)中BRD4表达;BRD4拮抗剂GSK525762A(500nmo L/L)处理喉鳞状细胞癌Hep2细胞,24、48和72 h后CCK-8检测细胞增殖;Trans-well细胞迁移实验和侵袭实验分别检测GSK525762A给药前后Hep2细胞迁移和侵袭能力;qRT-PCR和Western blot检测GSK525762A给药前后Hep2细胞BRD4、E-Cadherin、N-Cadherin和Twist蛋白表达。结果:喉鳞状细胞癌肿瘤组织中BRD4表达高于癌旁组织和正常卵巢组织(P0.05);GSK525762A给药处理后Hep2细胞增殖能力、迁移和侵袭能力及BRD4和上皮间质转化相关蛋白N-Cadherin和Twist表达均明显低于阴性对照组(P0.05),而E-Cadherin表达的表达明显升高(P0.05)。结论:BRD4可能通过抑制E-Cadherin的表达,增加N-Cadherin和Twist的表达,进而激活上皮间质转化,促进喉鳞状细胞癌转移。

Role and Mechanism of BRD4 in the Process of Focal Squamous Cell Carcinoma Metastasis

ABSTRACT Objective: To investigate the role and mechanism of BRD4 in the process of focal squamous cell carcinoma metastasis. Methods: Eighty-six surgical specimens of patients with laryngeal squamous cell carcinoma admitted from April 2016 to April 2018 were enrolled. qRT-PCR, Western blot and immunohistochemical staining were used to detect BRD4 expression in tumor tissues, paracancerous tissues and normal tissues; BRD4 antagonist GSK525762A (500 nmoL/L) was used to treat laryngeal squamous cell carcinoma Hep2 cells, 24, 48 and 72 After C, CCK-8 was used to detect cell proliferation; Trans-well cell migration assay and invasion assay were used to detect the migration and invasion of Hep2 cells before and after GSK525762A administration; qRT-PCR and Western blot were used to detect E-Cadherin, N-Cadherin and Twist in Hep2 cells before and after GSK525762A administration. Results: The expression of BRD4 in laryngeal squamous cell carcinoma was higher than that in adjacent normal tissues and normal ovarian tissues (P<0.05). The proliferation, migration and invasion of Hep2 cells and the protein associated with epithelial-mesenchymal transition were regulated by GSK525762A. The expression of Cadherin and Twist was significantly lower than that of the negative control group (P<0.05), while the expression of E-Cadherin was significantly increased (P<0.05). Conclusion: BRD4 may increase the expression of N-Cadherin and Twist by inhibiting the expression of E-Cadherin, thereby activating epithelial-mesenchymal transition and promoting the metastasis of laryngeal squamous cell carcinoma.