miR-214通过PTEN调控AKT信号通路抑制鼻咽癌细胞凋亡

韩继波; 黄茂凌; 申丽君; 陈始明; 陶泽璋

doi:10.3971/j.issn.1000-8578.2020.20.0149

miR-214通过PTEN调控AKT信号通路抑制鼻咽癌细胞凋亡

miR-214 Inhibits Apoptosis of Nasopharyngeal Carcinoma Cells by Activating AKT Signaling Pathway Through Targeting PTEN

摘要

摘要:
目的探讨miR-214通过PTEN调控下游AKT信号通路激活对鼻咽癌细胞凋亡的影响。
方法 miR-214抑制剂转染鼻咽癌5-8F和6-10B细胞。MTT法评估细胞增殖情况。Annexin-V/PI染色法检测两种鼻咽癌细胞凋亡情况。Western blot和real-time PCR检测miR-214抑制剂对PTEN基因转录和蛋白表达以及对AKT信号通路和凋亡相关蛋白表达水平的影响。检测shRNA沉默PTEN基因转录和蛋白表达后，miR-214抑制剂对鼻咽癌细胞凋亡影响。
结果 miR-214抑制剂可抑制鼻咽癌细胞生长，诱导5-8F和6-10B细胞凋亡。miR-214通过靶向3'-UTR区域调控PTEN基因转录和蛋白表达。抑制miR-214可促进PTEN的表达，抑制AKT信号通路激活，进而调控下游细胞周期和凋亡相关蛋白表达。沉默PTEN基因转录和蛋白表达可逆转miR-214抑制剂对鼻咽癌细胞AKT信号通路活性和凋亡的影响。
结论 miR-214可通过直接靶向PTEN基因转录促进AKT信号通路激活抑制鼻咽癌细胞凋亡。

Abstract:
Objective To investigate the effect of miR-214 on the apoptosis of nasopharyngeal carcinoma (NPC) cells by regulating the activation of AKT signaling pathway through targeting PTEN.
Methods The 5-8F and 6-10B NPC cells were transfected with miR-214 inhibitor. MTT assay was performed to assess cell proliferation. Annexin-V/PI staining assay was used to evaluate cell apoptosis. The effects of miR-214 inhibitor on the expression levels of PTEN, AKT signaling pathway and apoptosis-associated proteins were assessed by Western blot or real-time PCR assay. PTEN were knocked down using the corresponding shRNA to investigate the effect of miR-214 inhibitor on the apoptosis of NPC cells.
Results miR-214 inhibitor suppressed the growth and induced the apoptosis of 5-8F and 6-10B cells. miR-214 regulated the expression of PTEN through targeting the 3'-UTR. Inhibition of miR-214 promoted PTEN expression, inactivated AKT signaling pathway, and then regulated downstream cell cycle and apoptosis-associated proteins expression. Knockdown of PTEN reversed the effects of miR-214 inhibitor on AKT signaling and cell apoptosis.
Conclusion miR-214 could regulate the apoptosis of NPC cells through directly targeting PTEN and then activating AKT signaling pathway.

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