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LINC00174通过抑制miR-4500促进BZW2表达,增强喉乳头状瘤的增殖和凋亡逃避。

Promotion of BZW2 by LINC00174 through miR-4500 inhibition enhances proliferation and apoptosis evasion in laryngeal papilloma.

作者信息

Liu Jiajia, Yang Tao, Zhang Ying, Wang Shuhui

机构信息

Department of Otolaryngology-Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha, 410011 Hunan China.

出版信息

Cancer Cell Int. 2020 Sep 29;20:471. doi: 10.1186/s12935-020-01559-3. eCollection 2020.

Abstract

BACKGROUND

We aimed to explore the roles of basic leucine zipper and W2 domains (BZW) 2 in the human papillomavirus-infected laryngeal papillomatosis.

METHODS

In the present study, BZW 2 knockdown and overexpressed cell lines were constructed. CCK-8 and colony formation assays were used to determine cell proliferation. Caspase-3 activity and nucleosomes fragmentation assays were used to determine cell apoptosis. qRT-PCR and Western blot were employed to evaluate the mRNA and protein levels of target genes, respectively. Luciferase and biotin-coupled miRNA pulldown assays were used to examine the interactions between mRNA and mRNA.

RESULTS

We observed the levels of BZW2 were up-regulated in the laryngeal papilloma (LP) tissues as compared with adjacent tissues. The knockdown of BZW2 significantly inhibited cell proliferation and promoted cell apoptosis in the LP cells. Additionally, we identified the expressions of BZW2 negatively regulated by miR-4500. Luciferase and biotin-coupled miRNA pulldown assays demonstrated that LINC00174 competed with the BZW2 for binding with miR-4500. Moreover, the results showed that LINC00174/miR-4500/BZW2 axis regulated cell proliferation and apoptosis.

CONCLUSION

Our results demonstrated that the regulation of LINC00174/miR-4500/BZW2 axis might be used as an effective strategy for treatment of human papillomavirus-infected laryngeal papillomatosis.

摘要

背景

我们旨在探讨碱性亮氨酸拉链和W2结构域(BZW)2在人乳头瘤病毒感染的喉乳头状瘤病中的作用。

方法

在本研究中,构建了BZW 2基因敲低和过表达的细胞系。采用CCK-8和集落形成试验来测定细胞增殖。采用半胱天冬酶-3活性和核小体片段化试验来测定细胞凋亡。分别采用qRT-PCR和蛋白质印迹法来评估靶基因的mRNA和蛋白质水平。采用荧光素酶和生物素偶联的miRNA下拉试验来检测mRNA与mRNA之间的相互作用。

结果

我们观察到与相邻组织相比,喉乳头状瘤(LP)组织中BZW2的水平上调。敲低BZW2可显著抑制LP细胞的增殖并促进其凋亡。此外,我们发现BZW2的表达受到miR-4500的负调控。荧光素酶和生物素偶联的miRNA下拉试验表明,LINC00174与BZW2竞争与miR-4500的结合。此外,结果表明LINC00174/miR-4500/BZW2轴调节细胞增殖和凋亡。

结论

我们的结果表明,LINC00174/miR-4500/BZW2轴的调控可能是治疗人乳头瘤病毒感染的喉乳头状瘤病的有效策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3e6/7525952/c99abf34cbc0/12935_2020_1559_Fig1_HTML.jpg

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