CELF6 通过调控 FBP1 mRNA 的稳定性来调节三阴性乳腺癌的进展。

CELF6 modulates triple-negative breast cancer progression by regulating the stability of FBP1 mRNA.

机构信息

Department of Breast Surgery, Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, No. 218, Jixi Road, Hefei, 230022, Anhui, China.

Department of Thyroid and Breast Surgery, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, No. 1095, Jiefang Avenue, Wuhan, 430030, Hubei, China.

出版信息

Breast Cancer Res Treat. 2020 Aug;183(1):71-82. doi: 10.1007/s10549-020-05753-9. Epub 2020 Jun 29.

DOI:10.1007/s10549-020-05753-9

PMID:32601971

Abstract

BACKGROUND

Triple-negative breast cancer (TNBC) remains a great challenge in clinical treatment due to a shortage of effective therapeutic targets and acquired chemoresistance. Here, we identified the role of an RNA-binding protein, CUG-BP Elav-like family member 6 (CELF6), in the TNBC development and paclitaxel (PTX) chemoresistance.

METHODS

Stable CELF6-overexpressing cell lines were established in BT549 and MDA-MB-231 cells. Cell proliferation was determined using cell counting, two-dimensional colony formation, and MTT assay. Meanwhile, cell migration and cell invasion were detected by Transwell assay. Furthermore, the downstream target gene of CELF6 was identified and the direct interaction was further determined by luciferase reporter assay, immunoprecipitation, and RNA pull-down. Additionally, the PTX resistant cell line was established to determine the role of CELF6 in PTX resistance.

RESULTS

CELF6 overexpression suppressed cell proliferation, cell migration, and cell invasion. Mechanistically, Fructose-Bisphosphatase 1 (FBP1) was identified as the target gene of CELF6 and stabilized by CELF6 via binding 3'UTR. CELF6 overexpression mediated inhibition in TNBC development was dependent on FBP1. Moreover, CELF6 overexpression increased the sensitivity to PTX treatment.

CONCLUSION

CELF6 functions as a tumor suppressor by upregulating FBP 1 expression via stabilizing its mRNA, and thereby inhibits TNBC progression.

摘要

背景

由于缺乏有效的治疗靶点和获得性化疗耐药性，三阴性乳腺癌（TNBC）仍然是临床治疗的一大挑战。在这里，我们确定了 RNA 结合蛋白 CUG-BP Elav 样家族成员 6（CELF6）在 TNBC 发展和紫杉醇（PTX）化疗耐药性中的作用。

方法

在 BT549 和 MDA-MB-231 细胞中建立了稳定过表达 CELF6 的细胞系。通过细胞计数、二维集落形成和 MTT 测定来确定细胞增殖。同时，通过 Transwell 测定检测细胞迁移和细胞侵袭。此外，通过荧光素酶报告基因测定、免疫沉淀和 RNA 下拉确定 CELF6 的下游靶基因，并进一步确定其直接相互作用。此外，还建立了 PTX 耐药细胞系，以确定 CELF6 在 PTX 耐药性中的作用。

结果

CELF6 过表达抑制细胞增殖、细胞迁移和细胞侵袭。机制上，鉴定出 Fructose-Bisphosphatase 1（FBP1）是 CELF6 的靶基因，并通过与 3'UTR 结合由 CELF6 稳定。CELF6 过表达介导的 TNBC 发展抑制依赖于 FBP1。此外，CELF6 过表达增加了对 PTX 治疗的敏感性。