DNAJC12 通过激活 AKT 抑制阿霉素诱导的铁死亡和细胞凋亡，从而导致乳腺癌化疗耐药。

DNAJC12 causes breast cancer chemotherapy resistance by repressing doxorubicin-induced ferroptosis and apoptosis via activation of AKT.

机构信息

Department of Pathology, West China Hospital, Sichuan University, No. 37, Guo Xue Xiang, Chengdu, 610041, Sichuan, China; Institute of Clinical Pathology, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.

Institute of Clinical Pathology, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.

出版信息

Redox Biol. 2024 Apr;70:103035. doi: 10.1016/j.redox.2024.103035. Epub 2024 Jan 24.

DOI:10.1016/j.redox.2024.103035

PMID:38306757

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10847378/

Abstract

BACKGROUND

Chemotherapy is a primary treatment for breast cancer (BC), yet many patients develop resistance over time. This study aims to identify critical factors contributing to chemoresistance and their underlying molecular mechanisms, with a focus on reversing this resistance.

METHODS

We utilized samples from the Gene Expression Omnibus (GEO) and West China Hospital to identify and validate genes associated with chemoresistance. Functional studies were conducted using MDA-MB-231 and MCF-7 cell lines, involving gain-of-function and loss-of-function approaches. RNA sequencing (RNA-seq) identified potential mechanisms. We examined interactions between DNAJC12, HSP70, and AKT using co-immunoprecipitation (Co-IP) assays and established cell line-derived xenograft (CDX) models for in vivo validations.

RESULTS

Boruta analysis of four GEO datasets identified DNAJC12 as highly significant. Patients with high DNAJC12 expression showed an 8 % pathological complete response (pCR) rate, compared to 38 % in the low expression group. DNAJC12 inhibited doxorubicin (DOX)-induced cell death through both ferroptosis and apoptosis. Combining apoptosis and ferroptosis inhibitors completely reversed DOX resistance caused by DNAJC12 overexpression. RNA-seq suggested that DNAJC12 overexpression activated the PI3K-AKT pathway. Inhibition of AKT reversed the DOX resistance induced by DNAJC12, including reduced apoptosis and ferroptosis, restoration of cleaved caspase 3, and decreased GPX4 and SLC7A11 levels. Additionally, DNAJC12 was found to increase AKT phosphorylation in an HSP70-dependent manner, and inhibiting HSP70 also reversed the DOX resistance. In vivo studies confirmed that AKT inhibition reversed DNAJC12-induced DOX resistance in the CDX model.

CONCLUSION

DNAJC12 expression is closely linked to chemoresistance in BC. The DNAJC12-HSP70-AKT signaling axis is crucial in mediating resistance to chemotherapy by suppressing DOX-induced ferroptosis and apoptosis. Our findings suggest that targeting AKT and HSP70 activities may offer new therapeutic strategies to overcome chemoresistance in BC.

摘要

背景

化疗是乳腺癌（BC）的主要治疗方法，但许多患者随着时间的推移会产生耐药性。本研究旨在确定导致化疗耐药的关键因素及其潜在的分子机制，重点是逆转这种耐药性。

方法

我们利用来自基因表达综合数据库（GEO）和华西医院的样本，鉴定和验证与化疗耐药相关的基因。使用 MDA-MB-231 和 MCF-7 细胞系进行功能研究，包括功能获得和功能丧失方法。RNA 测序（RNA-seq）鉴定潜在机制。我们使用免疫共沉淀（Co-IP）检测 DNAJC12、HSP70 和 AKT 之间的相互作用，并建立细胞系衍生的异种移植（CDX）模型进行体内验证。

结果

对四个 GEO 数据集的 Boruta 分析表明 DNAJC12 高度显著。高 DNAJC12 表达的患者病理完全缓解（pCR）率为 8%，而低表达组为 38%。DNAJC12 通过铁死亡和细胞凋亡抑制多柔比星（DOX）诱导的细胞死亡。联合凋亡和铁死亡抑制剂可完全逆转 DNAJC12 过表达引起的 DOX 耐药性。RNA-seq 表明 DNAJC12 过表达激活了 PI3K-AKT 通路。AKT 抑制逆转了 DNAJC12 诱导的 DOX 耐药性，包括减少凋亡和铁死亡、恢复 cleaved caspase 3 和降低 GPX4 和 SLC7A11 水平。此外，发现 DNAJC12 以 HSP70 依赖的方式增加 AKT 磷酸化，抑制 HSP70 也逆转了 DOX 耐药性。体内研究证实 AKT 抑制在 CDX 模型中逆转了 DNAJC12 诱导的 DOX 耐药性。