外泌体 LOC85009 通过调节 ATG5 诱导的自噬抑制肺腺癌对多西他赛的耐药性。
Exosomal LOC85009 inhibits docetaxel resistance in lung adenocarcinoma through regulating ATG5-induced autophagy.
机构信息
Department of Medical Oncology, The First Affiliated Hospital of Soochow University, No.188 Shizi Street, Gusu District, Suzhou 215006, Jiangsu, China.
Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510062, Guangdong, China.
出版信息
Drug Resist Updat. 2023 Mar;67:100915. doi: 10.1016/j.drup.2022.100915. Epub 2022 Dec 26.
AIMS
This study aims at investigating the role of a neighbor long non-coding RNA (lncRNA) of HDAC4 (LOC85009) in docetaxel (DTX) resistance of lung adenocarcinoma (LUAD).
METHODS
RT-qPCR was used to analyze LOC85009 expression in DTX-resistant LUAD cells. In vitro and in vivo experiments were applied to detect the influence of LOC85009 on LUAD cell growth and xenograft tumor growth. DNA pull down assay, RNA pull down assay, ChIP assay, CoIP assay and RIP assay were performed to identify the direct interactions between factors.
RESULTS
LOC85009 was lowly-expressed in DTX-resistant LUAD cells. Functionally, LOC85009 overexpression inhibited DTX resistance and cell proliferation but triggered cell apoptosis. Moreover, we identified that LOC85009 was transferred from LUAD cells to DTX-resistant LUAD cells via exosomes. Exosomal LOC85009 inhibited DTX resistance, proliferation and autophagy while induced apoptosis in DTX-resistant cells. Additionally, we found that LOC85009 sequestered ubiquitin-specific proteinase 5 (USP5) to destabilize upstream transcription factor 1 (USF1) protein, thereby inactivating ATG5 transcription.
CONCLUSIONS
Exosomal LOC85009 inhibits DTX resistance through regulation of ATG5-induced autophagy via USP5/USF1 axis, suggesting that LOC85009 might be a potential target to reverse DTX resistance in the treatment of LUAD.
目的
本研究旨在探讨 HDAC4（LOC85009）的一个邻近长链非编码 RNA（lncRNA）在肺腺癌（LUAD）中多西紫杉醇（DTX）耐药中的作用。
方法
采用 RT-qPCR 分析 DTX 耐药 LUAD 细胞中 LOC85009 的表达。应用体外和体内实验检测 LOC85009 对 LUAD 细胞生长和异种移植肿瘤生长的影响。进行 DNA 下拉实验、RNA 下拉实验、ChIP 实验、CoIP 实验和 RIP 实验，以鉴定各因素之间的直接相互作用。
结果
LOC85009 在 DTX 耐药 LUAD 细胞中低表达。功能上，LOC85009 过表达抑制 DTX 耐药和细胞增殖，但触发细胞凋亡。此外，我们发现 LOC85009 通过外泌体从 LUAD 细胞转移到 DTX 耐药 LUAD 细胞。外泌体 LOC85009 抑制 DTX 耐药、增殖和自噬，同时诱导 DTX 耐药细胞凋亡。此外，我们发现 LOC85009 将泛素特异性蛋白酶 5（USP5）隔离以破坏上游转录因子 1（USF1）蛋白，从而使 ATG5 转录失活。
结论
外泌体 LOC85009 通过 USP5/USF1 轴调节 ATG5 诱导的自噬来抑制 DTX 耐药，表明 LOC85009 可能是逆转 LUAD 治疗中 DTX 耐药的潜在靶点。
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