FOXD3 通过 miR-335/DAAM1/肌球蛋白 II 轴依赖性机制赋予卵巢癌化疗敏感性。

FOXD3 confers chemo-sensitivity in ovarian cancer through a miR-335/DAAM1/myosin II axis-dependent mechanism.

机构信息

The First Affiliated Hospital, Department of Gynecology, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, China.

The First Affiliated Hospital, Reproductive Medicine Center, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, China.

出版信息

J Ovarian Res. 2023 Jan 10;16(1):8. doi: 10.1186/s13048-022-01077-y.

DOI:10.1186/s13048-022-01077-y

PMID:36627652

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9830800/

Abstract

BACKGROUND

Chemotherapy is among the most common treatment methods for ovarian cancer (OC). However, chemoresistance limits the effectiveness of chemotherapy and leads to treatment failure. We herein investigate the biological effect of forkhead box D3 (FOXD3) in the chemoresistance of OC cells.

METHODS

Expression of FOXD3, miR-335 and disheveled-associated activator of morphogenesis 1 (DAAM1) was detected in OC cells and tissues. The regulatory network of FOXD3/miR-335/DAAM1 was validated by dual-luciferase reporter and ChIP assays in vitro. After ectopic expression and depletion experiments in carboplatin/paclitaxel (CP)-resistant (A2780CP) or sensitive (A2780S) OC cells, cell viability, colony formation and apoptosis were tested by CCK-8 assay, colony formation assay and flow cytometry respectively. Effects of FOXD3 on the chemoresistance of OC cells in vivo were evaluated in OC xenografts in nude mice.

RESULTS

Overexpression of FOXD3 impaired the proliferation and chemoresistance of OC cells, which was related to the promotion of the miR-335 expression. Functionally, DAAM1 was a putative target of miR-335. Silencing of DAAM1 was responsible for the inhibition of myosin II activation, consequently leading to suppressed OC cell proliferation and chemoresistance. In vivo results further showed that FOXD3 weakened the chemoresistance of OC cells to CP.

CONCLUSION

Taken together, we unveil a novel FOXD3/miR-335/DAAM1/myosin II axis that regulates the chemoresistance of OC both in vitro and in vivo.

摘要

背景

化疗是卵巢癌（OC）最常见的治疗方法之一。然而，化疗耐药性限制了化疗的有效性，并导致治疗失败。我们在此研究叉头框 D3（FOXD3）在 OC 细胞化疗耐药中的生物学效应。

方法

检测 OC 细胞和组织中 FOXD3、miR-335 和形态发生中卷曲相关激活物 1（DAAM1）的表达。体外通过双荧光素酶报告和染色质免疫沉淀实验验证 FOXD3/miR-335/DAAM1 的调控网络。在卡铂/紫杉醇（CP）耐药（A2780CP）或敏感（A2780S）OC 细胞中外源表达和耗竭实验后，通过 CCK-8 assay、集落形成 assay 和流式细胞术分别检测细胞活力、集落形成和细胞凋亡。在裸鼠 OC 异种移植瘤中评估 FOXD3 对 OC 细胞化疗耐药性的体内影响。

结果

FOXD3 的过表达削弱了 OC 细胞的增殖和化疗耐药性，这与促进 miR-335 的表达有关。功能上，DAAM1 是 miR-335 的一个假定靶点。沉默 DAAM1 负责肌球蛋白 II 激活的抑制，进而导致 OC 细胞增殖和化疗耐药性的抑制。体内结果进一步表明，FOXD3 削弱了 OC 细胞对 CP 的化疗耐药性。