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miR-375 通过抑制 PDK1 抑制鼻咽癌细胞的增殖和侵袭。

miR-375 Inhibits the Proliferation and Invasion of Nasopharyngeal Carcinoma Cells by Suppressing PDK1.

机构信息

Department of Otolaryngology, The Third Affiliated Hospital of Wenzhou Medical University, Ruian, Zhejiang, China.

出版信息

Biomed Res Int. 2020 Mar 21;2020:9704245. doi: 10.1155/2020/9704245. eCollection 2020.

Abstract

PURPOSE

In patients with nasopharyngeal carcinoma (NPC), the expression of PDK1 is remarkably improved in NPC tissue and correlated with the clinicopathological severity of NPC. We expressed miR-375 in NPC cells to study the effects on PDK1 gene expression. We also investigated the mechanism by which miR-375 affects the biological behavior of NPC cells through effects on PDK1.

METHODS

qRT-PCR was carried out to analyze miR-375 and PDK1 levels in NPC cells. NPC cells were transfected with miR-375 inhibitor or miR-375 mimic. CCK-8 testing, colony formation testing, transwell testing, and flow cytometry analysis were carried out to quantify the cells' biological behavior. Rescue experiments demonstrated that the recovery of PDK1 expression was able to reverse the influence of miR-375 inhibition on NPC diffusion and intrusion. The interaction between miR-375 and PDK1 was verified by dual-luciferase reporter gene testing.

RESULTS

The results revealed that miR-375 has a negative regulatory effect on PDK1 expression in NPC cells. Furthermore, PDK1 is a target gene for miR-375. The empirical results obtained demonstrated a negative correlation between tumor development and the level of miR-375 expression in NPC tissues. The excessive expression of miR-375 and the downregulation of PDK1 facilitated the diffusion and invasion of NPC cells.

CONCLUSION

The diffusion and incursion of NPC cells may be inhibited by direct targeting of PDK1 and decreasing the expression of miR-375. Our study highlights efforts to target PDK1 and miR-375 as potential therapeutic strategies for use in the treatment of NPC.

摘要

目的

在鼻咽癌(NPC)患者中,PDK1 的表达在 NPC 组织中显著提高,并与 NPC 的临床病理严重程度相关。我们在 NPC 细胞中表达 miR-375,以研究其对 PDK1 基因表达的影响。我们还通过研究 miR-375 对 NPC 细胞生物学行为的影响来研究其影响 PDK1 的机制。

方法

采用 qRT-PCR 分析 NPC 细胞中 miR-375 和 PDK1 的水平。用 miR-375 抑制剂或 miR-375 模拟物转染 NPC 细胞。用 CCK-8 检测、集落形成检测、transwell 检测和流式细胞术分析来量化细胞的生物学行为。通过挽救实验证实,PDK1 表达的恢复能够逆转 miR-375 抑制对 NPC 扩散和浸润的影响。通过双荧光素酶报告基因检测验证 miR-375 和 PDK1 之间的相互作用。

结果

结果表明,miR-375 对 NPC 细胞中 PDK1 的表达具有负调控作用。此外,PDK1 是 miR-375 的靶基因。实验结果表明,miR-375 在 NPC 组织中的表达水平与肿瘤的发展呈负相关。miR-375 的过表达和 PDK1 的下调促进了 NPC 细胞的扩散和侵袭。

结论

通过直接靶向 PDK1 并降低 miR-375 的表达,可能抑制 NPC 细胞的扩散和侵袭。我们的研究强调了靶向 PDK1 和 miR-375 作为治疗 NPC 的潜在治疗策略的努力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/98c2/7125451/9fdd090e45d0/BMRI2020-9704245.001.jpg

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