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miR-214 通过靶向 WWOX 和 PTEN 介导鼻咽癌细胞的增殖和凋亡。

MiR-214 Mediates Cell Proliferation and Apoptosis of Nasopharyngeal Carcinoma Through Targeting Both WWOX and PTEN.

机构信息

Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People's Republic of China.

Research Institute of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, People's Republic of China.

出版信息

Cancer Biother Radiopharm. 2020 Oct;35(8):615-625. doi: 10.1089/cbr.2019.2978. Epub 2020 Feb 26.

Abstract

This study aimed to investigate interactions between miR-214, PTEN, and WWOX and their effect on AKT signaling during the NPC progression. Nasopharyngeal carcinoma (NPC) was highly prevalent with poor prognosis among the patients. MiR-214 reported as an important NPC biomarker was associated with regulation of biological functions. 5-8F and 6-10B NPC cells were transfected with miR-214 inhibitor. MTT and colony formation assays were performed to assess cell proliferation. PI staining assay was performed to determine distribution of cell cycle. Annexin-V/PI staining assay was used to evaluate cell apoptosis in NPC. The effects of miR-214 inhibitor on the expression levels of PTEN, WWOX, AKT signaling pathway, cell-cycle-, and apoptosis-associated proteins were assessed by Western blotting or qRT-PCR assay. and were knocked down using the corresponding shRNA to investigate their effects on miR-214 inhibitor involved in proapoptosis and antiproliferation mechanisms in NPC. Inhibition of miR-214 suppressed cell growth and induced apoptosis of 5-8F and 6-10B cells. MiR-214 regulated the expression of both and through targeting the 3'-UTR. Inhibition of miR-214 promoted WWOX and PTEN expression, inactivated AKT signaling pathway, and regulated cell-cycle- and apoptosis-associated proteins. Knockdown of or reversed effects of miR-214 inhibitor on AKT signaling, cell proliferation, and apoptosis. MiR-214 was suggested to induce cell proliferation and inhibit cell apoptosis of NPC through directly targeting both PTEN and WWOX, which provided a novel therapeutic target for clinical treatment of NPC.

摘要

本研究旨在探讨 miR-214、PTEN 和 WWOX 之间的相互作用及其对 NPC 进展过程中 AKT 信号通路的影响。鼻咽癌(NPC)是一种高发的恶性肿瘤,患者预后较差。miR-214 作为一种重要的 NPC 生物标志物,与调节生物学功能有关。我们用 miR-214 抑制剂转染 5-8F 和 6-10B NPC 细胞。用 MTT 和集落形成实验检测细胞增殖,PI 染色实验检测细胞周期分布,Annexin-V/PI 染色实验检测 NPC 细胞凋亡。用 Western blot 或 qRT-PCR 实验检测 miR-214 抑制剂对 PTEN、WWOX、AKT 信号通路、细胞周期和凋亡相关蛋白表达水平的影响。用相应的 shRNA 敲低 和 ,研究它们对 miR-214 抑制剂在 NPC 中促凋亡和抗增殖机制中的作用。抑制 miR-214 抑制了 5-8F 和 6-10B 细胞的生长并诱导其凋亡。miR-214 通过靶向 3'-UTR 调节 和 的表达。抑制 miR-214 促进了 WWOX 和 PTEN 的表达,使 AKT 信号通路失活,并调节细胞周期和凋亡相关蛋白。敲低 或 逆转了 miR-214 抑制剂对 AKT 信号、细胞增殖和凋亡的影响。miR-214 可能通过直接靶向 PTEN 和 WWOX 诱导 NPC 细胞增殖和抑制细胞凋亡,为 NPC 的临床治疗提供了新的治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0843/7578184/be469d5d003d/cbr.2019.2978_figure1.jpg

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