SP1 激活的 LINC01614 通过 WNT/β-catenin 信号通路促进三阴性乳腺癌细胞的恶性行为。

LINC01614 activated by SP1 promoted malignant behavior of triple-negative breast cancer cells via the WNT/b-Catenin signaling pathway.

机构信息

Department of Breast and Thyroid Surgery, Renmin Hospital of Wuhan University, Wuhan, China.

Department of Oncology, Yichang Central People's Hospital, Yichang, China.

出版信息

Rev Invest Clin. 2024 Oct 17;76(4):185-194. doi: 10.24875/RIC.24000093.

DOI:10.24875/RIC.24000093

PMID:39419019

Abstract

BACKGROUND

Triple-negative breast cancer (TNBC) represents the most aggressive subtype of breast cancer (BC), characterized by a dismal prognosis. Dysregulated long non-coding RNA LINC01614 might be a potential biomarker for BC as previously reported. Nevertheless, its functions and mechanism in TNBC cells are unclear.

OBJECTIVES

The study aimed to study the effects of LINC01614 on TNBC cell migration, invasion, and epithelial-mesenchymal transition (EMT) process as well as the related mechanism.

METHODS

Reverse transcription quantitative polymerase chain reaction was performed to detect the expression of LINC01614 and SP1 in TNBC cells and tissues. The cellular localization of LINC01614 was determined by subcellular fraction assays. Cell counting kit-8 and Transwell invasion assays were conducted for measurement of TNBC cell viability and invasive ability. Cell migration was performed using wound healing assays and Transwell migration assays. Chromatin immunoprecipitation assays and luciferase reporter assays were used to explore the interaction between SP1 and LINC01614. Western blotting was used to assess protein levels of factors involved in EMT process and Wnt/ß-catenin signaling in TNBC cells.

RESULTS

LINC01614 expression was elevated in TNBC tissues and cells. LINC01614 knockdown inhibited cell viability as well as migratory and invasive abilities of TNBC cells. LINC01614 knockdown also obstructed EMT process, as shown by E-cadherin upregulation and vimentin downregulation in TNBC cells. SP1 directly bound to the promoter of LINC01614 and activated LINC01614 expression. SP1 overexpression reversed the suppressive effect of LINC01614 knockdown on TNBC cell migration, invasion, and EMT process. Protein levels of Wnt and ß-catenin were diminished by LINC01614 knockdown, and the trend was partially rescued by SP1 overexpression.

CONCLUSION

SP1-induced LINC01614 promoted malignant behavior of TNBC cells by activating the Wnt/ß-catenin signaling pathway.

摘要

背景

三阴性乳腺癌（TNBC）是乳腺癌（BC）中最具侵袭性的亚型，预后不良。先前有报道称，失调的长非编码 RNA LINC01614 可能是 BC 的潜在生物标志物。然而，其在 TNBC 细胞中的功能和机制尚不清楚。

目的

本研究旨在研究 LINC01614 对 TNBC 细胞迁移、侵袭和上皮-间充质转化（EMT）过程的影响及其相关机制。

方法

采用逆转录定量聚合酶链反应检测 TNBC 细胞和组织中 LINC01614 和 SP1 的表达。通过亚细胞分离试验确定 LINC01614 的细胞定位。细胞计数试剂盒-8 法和 Transwell 侵袭实验用于测量 TNBC 细胞活力和侵袭能力。采用划痕愈合实验和 Transwell 迁移实验检测细胞迁移。染色质免疫沉淀实验和荧光素酶报告基因实验用于探索 SP1 与 LINC01614 之间的相互作用。Western blot 法检测 TNBC 细胞中 EMT 过程和 Wnt/β-catenin 信号通路相关因子的蛋白水平。

结果

LINC01614 在 TNBC 组织和细胞中表达上调。LINC01614 敲低抑制 TNBC 细胞的活力以及迁移和侵袭能力。LINC01614 敲低还阻止了 EMT 过程，表现为 TNBC 细胞中 E-钙黏蛋白上调和波形蛋白下调。SP1 直接结合到 LINC01614 的启动子上并激活 LINC01614 的表达。SP1 过表达逆转了 LINC01614 敲低对 TNBC 细胞迁移、侵袭和 EMT 过程的抑制作用。LINC01614 敲低降低了 Wnt 和 β-连环蛋白的蛋白水平，SP1 过表达部分挽救了这一趋势。