Department of Radiation Oncology, Huizhou Municipal Central Hospital, Guangdong, China.
Bioengineered. 2021 Dec;12(1):7325-7334. doi: 10.1080/21655979.2021.1979356.
MicroRNAs (miRNAs) are revealed to participate in the progression of multiple malignancies, including nasopharyngeal carcinoma (NPC). This work is intended to decipher the function of microRNA-195-3p (miR-195-3p) in regulating the radiosensitivity of NPC cells and its mechanism. MiR-195-3p and cyclin-dependent kinase 1 (CDK1) expressions were detected in NPC tissues and cells using qRT-PCR and Western blot, respectively. Moreover, radiation-resistant cell lines were induced by continuous irradiation with different doses. Furthermore, the CCK-8 experiment, colony formation assay and flow cytometry were utilized to examine the growth, apoptosis and cell cycle of radioresistant cells. Bioinformatics prediction and dual-luciferase reporter gene assay were applied to prove the targeting relationship between miR-195-3p and CDK1 mRNA 3'UTR. The data showed that miR-195-3p was remarkably down-modulated in NPC tissues and was associated with increased tumor grade, lymph node metastasis and clinical stage of the patients. MiR-195-3p expression was significantly down-modulated in radiation-resistant NPC tissues and NPC cell lines relative to radiation-sensitive NPC tissues and human nasopharyngeal epithelial cells, while CDK1 expression was notably up-modulated. MiR-195-3p overexpression inhibited the growth of NPC cells, decreased radioresistance, promoted apoptosis, and impeded the cell cycle progression. CDK1 was a target gene of miR-195-3p, and CDK1 overexpression counteracted the effects of miR-195-3p on NPC cell growth, apoptosis, cell cycle progression and radiosensitivity. In summary, miR-195-3p improves the radiosensitivity of NPC cells by targeting and regulating CDK1.
微小 RNA（miRNA）被揭示参与多种恶性肿瘤的进展，包括鼻咽癌（NPC）。本工作旨在破译 microRNA-195-3p（miR-195-3p）在调节 NPC 细胞放射敏感性及其机制中的作用。使用 qRT-PCR 和 Western blot 分别检测 NPC 组织和细胞中的 miR-195-3p 和细胞周期蛋白依赖性激酶 1（CDK1）表达。此外，通过用不同剂量连续照射诱导耐辐射细胞系。进一步，通过 CCK-8 实验、集落形成实验和流式细胞术检测耐辐射细胞的生长、凋亡和细胞周期。生物信息学预测和双荧光素酶报告基因实验用于证明 miR-195-3p 与 CDK1 mRNA 3'UTR 之间的靶向关系。数据表明，miR-195-3p 在 NPC 组织中显著下调，与患者肿瘤分级增加、淋巴结转移和临床分期相关。miR-195-3p 在耐辐射 NPC 组织和 NPC 细胞系中的表达明显下调，而 CDK1 的表达明显上调。miR-195-3p 过表达抑制 NPC 细胞生长，降低耐辐射性，促进细胞凋亡，抑制细胞周期进程。CDK1 是 miR-195-3p 的靶基因，CDK1 过表达抵消了 miR-195-3p 对 NPC 细胞生长、凋亡、细胞周期进程和放射敏感性的影响。总之，miR-195-3p 通过靶向和调节 CDK1 提高 NPC 细胞的放射敏感性。
