Department of Gynecology, Shulan (Hangzhou) Hospital, Shulan International Medical College, Zhejiang Shuren University, Hangzhou, China.
Adv Clin Exp Med. 2024 Mar;33(3):273-282. doi: 10.17219/acem/168342.
Cervical cancer is prevalent throughout the world, and microRNA-497-5p (miR-497-5p) plays an important role in its development. However, the specific mechanism by which miR-497-5p targets the transferrin receptor (TFRC) during cervical cancer development has not been clarified.
The aim of the study was to unravel TFRC expression and its role in cervical cancer cells, as well as the impact of the miR-497-5p/TFRC axis on cervical cancer cells.
The target mRNA was determined through differential analysis, followed by the evaluation of its impact on survival and clinical staging. Then, quantitative real-time polymerase chain reaction (qPCR) was conducted to analyze the TFRC mRNA level in cervical cancer cells and normal cervical epithelial cells. Western blot (WB) was utilized to examine the expression levels of TFRC, cleaved caspase-3, cleaved caspase-9, and epithelial-mesenchymal transition (EMT)-related proteins. The miRNAs upstream of the target mRNA were predicted, and Pearson correlation analysis was performed, followed by the validation through the dual-luciferase reporter assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were performed to analyze cancer cell viability, followed by a transwell assay aimed at measuring cell migratory and invasive abilities. Finally, flow cytometry was conducted to examine cell apoptosis and cell cycle.
The transferrin receptor was significantly increased in cervical cancer cells and positively associated with clinical T and N stages. Silencing TFRC could constrain cell proliferative, migratory and invasive abilities, arrest the cell cycle and facilitate cell apoptosis in cervical cancer cells. The bioinformatics analysis showed a significantly negative correlation between miR-497-5p and TFRC in cervical cancer. Moreover, upregulated miR-497-5p hampered cervical cancer progression and decreased TFRC expression. The overexpression of TFRC reversed the suppressive impact of miR-497-5p overexpression on cervical cancer progression.
The modulatory role of the miR-497-5p/TFRC axis was confirmed in cervical cancer cells. This axis may present a new avenue for the diagnosis of cervical cancer and provide a novel target for cervical cancer treatment.
宫颈癌在全球范围内普遍存在，miR-497-5p 在其发展过程中起着重要作用。然而，miR-497-5p 靶向转铁蛋白受体（TFRC）在宫颈癌发展过程中的具体机制尚不清楚。
本研究旨在阐明 TFRC 的表达及其在宫颈癌细胞中的作用，以及 miR-497-5p/TFRC 轴对宫颈癌细胞的影响。
通过差异分析确定靶 mRNA，然后评估其对生存和临床分期的影响。然后，通过定量实时聚合酶链反应（qPCR）分析宫颈癌细胞和正常宫颈上皮细胞中 TFRC mRNA 水平。Western blot（WB）检测 TFRC、裂解的半胱天冬酶-3、裂解的半胱天冬酶-9 和上皮-间充质转化（EMT）相关蛋白的表达水平。预测靶 mRNA 的上游 miRNA，并进行 Pearson 相关性分析，然后通过双荧光素酶报告基因实验进行验证。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）和集落形成实验分析癌细胞活力，然后通过 Transwell 实验测量细胞迁移和侵袭能力。最后，通过流式细胞术检测细胞凋亡和细胞周期。
转铁蛋白受体在宫颈癌细胞中显著增加，并与临床 T 和 N 分期呈正相关。沉默 TFRC 可抑制宫颈癌细胞的增殖、迁移和侵袭能力，阻滞细胞周期并促进细胞凋亡。生物信息学分析显示 miR-497-5p 与宫颈癌中的 TFRC 呈显著负相关。此外，上调 miR-497-5p 可阻碍宫颈癌的进展并降低 TFRC 表达。TFRC 的过表达逆转了 miR-497-5p 过表达对宫颈癌进展的抑制作用。
证实了 miR-497-5p/TFRC 轴在宫颈癌细胞中的调节作用。该轴可能为宫颈癌的诊断提供新途径，并为宫颈癌的治疗提供新靶点。
