Department of Dermatology, The Central hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan City, 430014, Hubei Province, China.
Department of Gastroenterology, Ganzhou People's Hospita, Ganzhou City, 341000, Jiangxi Province, China.
Pathol Oncol Res. 2020 Jul;26(3):1851-1859. doi: 10.1007/s12253-019-00782-2. Epub 2019 Nov 28.
Condyloma acuminate (CA) is a communicable disease caused by human papillomavirus (HPV). This study aimed to study the targeting relationship between miR-34a-5p and Jagged 1 (JAG1), as well as its regulatory effect in HPV-infected cells. Human keratinocyte HaCaT cells were infected with HPV16E6, and CA tissues were collected. The expression level of miR-34a-5p and JAG1 were detected in CA tissues and HPV-HaCaT cells. Cell proliferation, migration and invasion were respectively measured using 3-(4, 5)-dimethylthiahiazo-(-z-y1)-3, 5-diphenytetrazoliumromide (MTT), cell wound healing and Transwell assay. The potential binding sites of miR-34a-5p and JAG1 were predicted by website TargetScan, and confirmed using dual luciferase reporter gene assay. The proteins of Notch1 pathway-related were assessed using western blotting. The results showed that miR-34a-5p expression was decreased, and JAG1 expression was increased in CA tissues and HPV-HaCaT cells. Cell proliferation, migration and invasion were decreased when miR-34a-5p over-expression and JAG1 knock-down in HPV-HaCaT cells. Furthermore, miR-34a-5p had a targeting effect on JAG1. The expression level of Notch1, NICD, Hes1 and Hey1 were increased when miR-34a-5p knock-down. miR-34a-5p could inhibit cell development, and regulate the activity of Notch1 pathway through targeting JAG1 expression in HPV-infected keratinocytes.
尖锐湿疣(CA)是一种由人乳头瘤病毒(HPV)引起的传染病。本研究旨在研究 miR-34a-5p 与 Jagged 1(JAG1)的靶向关系及其在 HPV 感染细胞中的调节作用。用人乳头瘤病毒 16E6 感染人角质形成细胞 HaCaT 细胞,并收集 CA 组织。检测 CA 组织和 HPV-HaCaT 细胞中 miR-34a-5p 和 JAG1 的表达水平。分别用 3-(4,5)-二甲基噻唑-(-z-y1)-3,5-二苯基四氮唑溴盐(MTT)、细胞划痕愈合和 Transwell 实验检测细胞增殖、迁移和侵袭。通过网站 TargetScan 预测 miR-34a-5p 和 JAG1 的潜在结合位点,并通过双荧光素酶报告基因实验进行验证。用 Western blot 检测 Notch1 通路相关蛋白。结果显示,CA 组织和 HPV-HaCaT 细胞中 miR-34a-5p 表达降低,JAG1 表达升高。miR-34a-5p 过表达和 JAG1 敲低时,HPV-HaCaT 细胞的细胞增殖、迁移和侵袭能力降低。此外,miR-34a-5p 对 JAG1 有靶向作用。miR-34a-5p 敲低时,Notch1、NICD、Hes1 和 Hey1 的表达水平升高。miR-34a-5p 可通过靶向 JAG1 表达抑制 HPV 感染角质形成细胞的发育,并调节 Notch1 通路活性。