HLTF/SERPINE1轴在宫颈癌前病变向宫颈癌的体外进展过程中发挥着至关重要的促癌作用

IF 2 4区 医学 Q2 OBSTETRICS & GYNECOLOGY
Yong Wang, Yudi Tan, Shasha Yang, Jinkong Wei, Yuying Wei, Junying Chen
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引用次数: 0

摘要

目的:宫颈癌(CC)在全球妇女中普遍存在,而且发病风险越来越高。寻找治疗宫颈癌的有效方法至关重要。本研究旨在探讨 SERPINE1 对宫颈癌前病变发展为 CC 的影响:设计:转录组测序和体外细胞系研究。参与者/材料:宫颈癌前病变和CC样本、人宫颈上皮永生细胞系H8、人CC细胞系HeLa和CaSki:应用新一代测序技术从宫颈癌前病变和CC样本中鉴定出100个差异表达基因(DEGs)。应用检索相互作用基因的搜索工具(STRING)数据库,我们进行了蛋白质-蛋白质相互作用(PPI)网络分析,从而筛选出在CC细胞中显著上调的丝氨酸蛋白酶抑制剂E族成员1(SERPINE1)。利用人类转录因子数据库(HumanTFDB)预测了螺旋酶样转录因子(HLTF)作为上游转录因子。染色质免疫共沉淀(ChIP)实验验证了SERPINE1与HLTF之间的相互作用。免疫组织化学(IHC)检测了SERPINE1和HLTF在CC组织中的表达。在上调或下调 SERPINE1 和 HLTF 后,进行实时定量反转录聚合酶链反应(qRT-PCR)以评估细胞中 SERPINE1 和 HLTF 的 mRNA 表达水平。细胞活力、迁移和侵袭分别采用 MTT 试验、细胞划痕试验和 Transwell 试验进行评估。Western印迹(WB)分析评估了基质金属蛋白酶和上皮-间质转化(EMT)相关蛋白表达水平的变化:结果:ChIP实验证实了HLTF和SERPINE1之间的相互作用。HLTF和SERPINE1在CC组织和细胞中上调,沉默SERPINE1可抑制EMT过程和CC细胞的活力、迁移和侵袭。然而,在CC细胞中过表达SERPINE1则表现出相反的趋势。拯救实验表明,沉默HLTF抑制了CC细胞的活力、迁移和侵袭,而过表达SERPINE1则可恢复活力、迁移和侵袭:局限性:HLTF/SERPINE1轴对CC恶性进展的影响尚未得到体内实验的证实:结论:HLTF可转录激活SERPINE1,促进宫颈癌前病变向CC发展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
HLTF/SERPINE1 Axis Plays a Crucial Pro-Oncogenic Role in the Progression from Cervical Precancerous Lesions to Cervical Carcinoma in vitro.

Objectives: Cervical carcinoma (CC) is prevalent among women worldwide with increasing risk. Finding effective methods for treating CC is of utmost importance. The aim of this study was to investigate the effect of SERPINE1 on the progression of cervical precancerous lesions to CC.

Design: This study used transcriptome sequencing and in vitro cell line. Participants/Materials: Cervical precancerous lesions and CC samples and human cervical epithelial immortalized cell line H8, human CC cell lines HeLa, and CaSki were involved in this study.

Setting and methods: Next-generation sequencing was applied to identify 100 differentially expressed genes from cervical precancerous lesions and CC samples. With the application of the Search Tool for the Retrieval of Interacting Genes (STRING) database, we carried out the protein-protein interaction network analysis, thus screening out serine protease inhibitor clade E member 1 (SERPINE1) with significant upregulation in CC cells. The helicase-like transcription factor (HLTF) was predicted as the upstream transcription factor using Human Transcription Factor Database (HumanTFDB). The chromatin immunoprecipitation (ChIP) experiment was conducted to validate the interaction between SERPINE1 and HLTF. The immunohistochemistry was employed to determine the expression of SERPINE1 and HLTF in CC tissues. Following the upregulation or downregulation of SERPINE1 and HLTF, the real-time quantitative reverse transcription polymerase chain reaction was carried out to assess mRNA expression levels of SERPINE1 and HLTF in cells. Cell viability, migration, and invasion were evaluated using MTT assay, cell scratch assay, and Transwell assay, respectively. Western blot analysis was conducted to assess changes in the expression levels of matrix metalloproteinases and proteins related to epithelial-mesenchymal transition (EMT).

Results: The ChIP experiment confirmed the interaction between HLTF and SERPINE1. HLTF and SERPINE1 were upregulated in CC tissues and cells, and silencing SERPINE1 inhibited the EMT process and viability, migration, and invasion of CC cells. However, overexpression of SERPINE1 in CC cells showed the opposite trend. Rescue experiments demonstrated that silencing HLTF repressed CC cell viability, migration, and invasion, which could be restored by overexpressing SERPINE1.

Limitations: The effect of the HLTF/SERPINE1 axis on CC malignant progression has not been confirmed by in vivo experiments.

Conclusion: HLTF transcriptionally activates SERPINE1, promoting the progression from cervical precancerous lesions to CC.

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来源期刊
CiteScore
4.20
自引率
4.80%
发文量
44
审稿时长
6-12 weeks
期刊介绍: This journal covers the most active and promising areas of current research in gynecology and obstetrics. Invited, well-referenced reviews by noted experts keep readers in touch with the general framework and direction of international study. Original papers report selected experimental and clinical investigations in all fields related to gynecology, obstetrics and reproduction. Short communications are published to allow immediate discussion of new data. The international and interdisciplinary character of this periodical provides an avenue to less accessible sources and to worldwide research for investigators and practitioners.
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