tRNA 衍生的片段 3'tRF-AlaAGC 通过与乳腺癌中的 TRADD 结合调节细胞化疗耐药性和 M2 巨噬细胞极化。
tRNA-derived fragment 3'tRF-AlaAGC modulates cell chemoresistance and M2 macrophage polarization via binding to TRADD in breast cancer.
机构信息
Department of Clinical Laboratory, Nanjing Medical University Affiliated Cancer Hospital & Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research, Baizi Ting No.42, Nanjing, 210009, China.
Jiangsu Key Laboratory of Molecular and Translational Cancer Research, Baizi Ting No.42, Nanjing, 210009, China.
出版信息
J Transl Med. 2024 Jul 30;22(1):706. doi: 10.1186/s12967-024-05513-z.
BACKGROUND
Drug resistance, including Adriamycin-based therapeutic resistance, remains a challenge in breast cancer (BC) treatment. Studies have revealed that macrophages could play a pivotal role in mediating the chemoresistance of cancer cells. Accumulating evidence suggests that tRNA-Derived small RNAs (tDRs) are associated the physiological and pathological processes in multiple cancers. However, the underlying mechanisms of tDRs on chemoresistance of BC in tumor-associated macrophages remain largely unknown.
METHODS
The high-throughput sequencing technique was used to screen tDRs expression profile in BC cells. Gain- and loss-of-function experiments and xenograft models were performed to verify the biological function of 3'tRF-Ala-AGC in BC cells. The CIBERSORT algorithm was used to investigate immune cell infiltration in BC tissues. To explore the role of 3'tRF-Ala-AGC in macrophages, M2 macrophages transfected with 3'tRF-Ala-AGC mimic or inhibitor were co-cultured with BC cells. Effects on Nuclear factor-κb (NF-κb) pathway were investigated by NF-κb nuclear translocation assay and western blot analysis. RNA pull-down assay was performed to identify 3'tRF-Ala-AGC interacting proteins.
RESULTS
A 3'tRF fragment of 3'tRF-AlaAGC was screened, which is significantly overexpressed in BC specimens and Adriamycin-resistant cells. 3'tRF-AlaAGC could promote cell malignant activity and facilitate M2 polarization of macrophages in vitro and in vivo. Higher expression of M2 macrophages were more likely to have lymph node metastasis and deeper invasion in BC patients. Mechanistically, 3'tRF-AlaAGC binds Type 1-associated death domain protein (TRADD) in BC cells, and suppression of TRADD partially abolished the enhanced effect of 3'tRF-AlaAGC mimic on phenotype of M2. The NF-κb signaling pathway was activated in BC cells co-cultured with M2 macrophages transfected with 3'tRF-AlaAGC mimic.
CONCLUSIONS
3'tRF-AlaAGC might modulate macrophage polarization via binding to TRADD and increase the effect of M2 on promoting the chemoresistance in BC cells through NF-κb signaling pathway.
背景
包括阿霉素治疗耐药在内的耐药性仍然是乳腺癌（BC）治疗的一个挑战。研究表明，巨噬细胞在介导癌细胞的化疗耐药中可能发挥关键作用。越来越多的证据表明 tRNA 衍生的小 RNA（tDRs）与多种癌症的生理和病理过程有关。然而，tDRs 对肿瘤相关巨噬细胞中 BC 化疗耐药的潜在机制在很大程度上尚不清楚。
方法
使用高通量测序技术筛选 BC 细胞中 tDRs 的表达谱。通过增益和失能实验以及异种移植模型验证 3'tRF-Ala-AGC 在 BC 细胞中的生物学功能。使用 CIBERSORT 算法研究 BC 组织中的免疫细胞浸润。为了探讨 3'tRF-Ala-AGC 在巨噬细胞中的作用，用 3'tRF-Ala-AGC 模拟物或抑制剂转染 M2 巨噬细胞，与 BC 细胞共培养。通过 NF-κb 核易位测定和 Western blot 分析研究 NF-κb 通路的作用。通过 RNA 下拉实验鉴定 3'tRF-Ala-AGC 的相互作用蛋白。
结果
筛选出 3'tRF-AlaAGC 的 3'tRF 片段，其在 BC 标本和阿霉素耐药细胞中显著过表达。3'tRF-AlaAGC 可促进细胞恶性活性，并促进体外和体内巨噬细胞的 M2 极化。BC 患者中 M2 巨噬细胞表达较高时，更有可能发生淋巴结转移和更深的浸润。机制上，3'tRF-AlaAGC 在 BC 细胞中与 1 型相关死亡域蛋白（TRADD）结合，抑制 TRADD 部分消除 3'tRF-AlaAGC 模拟物对 M2 表型增强的作用。共培养转染 3'tRF-AlaAGC 模拟物的 M2 巨噬细胞的 BC 细胞中 NF-κb 信号通路被激活。
结论
3'tRF-AlaAGC 可能通过与 TRADD 结合调节巨噬细胞极化，并通过 NF-κb 信号通路增加 M2 对促进 BC 细胞化疗耐药的作用。
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