miR-499b-5p 通过靶向 Notch1 信号通路抑制宫颈癌细胞增殖并诱导细胞凋亡。

MiR-499b-5p inhibits cervical cancer cell proliferation and induces apoptosis by targeting the Notch1 signaling pathway.

机构信息

Department of Gynaecology, First Affiliated Hospital, School of Medicine, Shihezi University, Xinjiang Uygur Autonomous Region, China.

出版信息

Eur Rev Med Pharmacol Sci. 2021 Oct;25(20):6220-6231. doi: 10.26355/eurrev_202110_26992.

DOI:10.26355/eurrev_202110_26992

PMID:34730202

Abstract

OBJECTIVE

We investigated the effect of miR-499b-5p on the tumorigenesis and development of cervical cancer by targeting the Notch1 signaling pathway to identify a new potential clinical target of cervical cancer.

PATIENTS AND METHODS

Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to determine the mRNA expression levels of Notch1 and miR-499b-5p in cervical cancer tissues/cell lines. Cell counting kit-8 (CCK-8) assay, transwell assay, and flow cytometry were conducted to detect cell viability, cell migration, and cell apoptosis abilities. A Dual-Luciferase reporter assay was performed to test the binding site between miR-499b-5p and Notch1. An in vivo experiment was carried out using nude mice, and xenograft tumor models were established.

RESULTS

OD450 of the SiHa and HeLa cells of the miR-499b-5p agomir group was lower than that of the miR-499b-5p agomir-NC group. More apoptotic cells and fewer invasive cells were found in the former than in the latter. MiR-499b-5p inhibited the viability and migration of cervical cancer cells and promoted their apoptosis. Further detection of the Luciferase reporter gene confirmed the binding site of miR-499b-5p to Notch1. Western blot results showed that miR-499b-5p inhibited the expression of Notch1 and activated the expression of ChK2 and p-p38MAPK. Notch1 knockdown also inhibited the viability and migration of cervical cancer cells and promoted their apoptosis. MiR-499b-5p overexpression prevented the tumorigenesis and development of cervical cancer in xenograft tumor models.

CONCLUSIONS

MiR-499b-5p inhibits the proliferation of cervical cancer cells and induces their apoptosis by targeting the Notch1 signaling pathway.

摘要

目的

通过靶向 Notch1 信号通路研究 miR-499b-5p 对宫颈癌发生发展的影响，寻找宫颈癌新的潜在临床治疗靶点。

方法

采用实时定量聚合酶链反应（qRT-PCR）检测宫颈癌组织/细胞中 Notch1 和 miR-499b-5p 的 mRNA 表达水平。细胞计数试剂盒（CCK-8）检测、Transwell 检测和流式细胞术检测细胞活力、细胞迁移和细胞凋亡能力。双荧光素酶报告基因实验检测 miR-499b-5p 与 Notch1 的结合位点。建立裸鼠体内实验，建立异种移植瘤模型。

结果

miR-499b-5p 激动剂组 SiHa 和 HeLa 细胞的 OD450 值低于 miR-499b-5p 激动剂-NC 组。前者凋亡细胞较多，侵袭细胞较少。miR-499b-5p 抑制宫颈癌细胞的活力和迁移，促进其凋亡。进一步检测荧光素酶报告基因证实了 miR-499b-5p 与 Notch1 的结合位点。Western blot 结果表明，miR-499b-5p 抑制 Notch1 的表达，激活 ChK2 和 p-p38MAPK 的表达。Notch1 敲低也抑制宫颈癌细胞的活力和迁移，促进其凋亡。miR-499b-5p 过表达可预防异种移植瘤模型中宫颈癌的发生发展。