环状 RNA SLC4A7 在耐药细胞衍生的外泌体中通过 miR-1205/MAPT 轴促进前列腺癌多西他赛耐药。

CircSLC4A7 in resistant-cells-derived exosomes promotes docetaxel resistance via the miR-1205/MAPT axis in prostate cancer.

机构信息

Department of Endocrinology, Jiangxi Provincial People's Hospital, The First Affiliated Hospital of Nanchang Medical College, Nanchang, China.

Department of Oncology, The First Affiliated Hospital of Nanchang University, Nanchang, China.

出版信息

IUBMB Life. 2024 Dec;76(12):1342-1355. doi: 10.1002/iub.2915. Epub 2024 Sep 12.

DOI:10.1002/iub.2915

PMID:39266461

Abstract

Prostate cancer (PCa) is a high-mortality cancer. Docetaxel (DCT) combined with second-generation anti-androgens is considered the golden standard therapy for PCa, whose application is limited for DCT resistance (DR). Therefore, exploring the mechanism of DR is of great importance. In this study, PCa cell lines of PC3 and DU145 were employed, and DR cells were constructed by treatment with graded DCT. CircSLC4A7, miR-1205, and microtubule-associated protein tau (MAPT) transfections were established. Cell counting kit-8 assay was performed to evaluate the cell activity and IC of DCT. After being treated with DCT, DR was assessed by colony formation assay, flow cytometry analysis, and terminal transferase-mediated UTP nick end-labeling assay. Real-time quantitative PCR and western blotting analysis evaluated the expression levels of genes. The dual-luciferase reporter gene assay verified the miR-1205 binding sites with circSLC4A7 and MAPT. An animal experiment was performed to assess the tumor growth influenced by circSLC4A7. After conducting DR cells and isolated exosomes, we found that not only co-culture with DR cells but also treatment with DR cells' exosomes would promote the DR of normal cells. Moreover, circSLC4A7 was highly expressed in DR cells and their exosomes. CircSLC4A7 overexpression enhanced DR, represented as raised IC50 of DCT, increased colony formation, and decreased cell apoptosis after DCT treatment, while circSLC4A7 knockdown had the opposite effect. MiR-1205 was confirmed as a circSLC4A7-sponged miRNA and miR-1205 inhibitor reversed the effect of sh-circSLC4A7. MAPT was further identified as a target of miR-1205 and had a similar effect with circSLC4A7. The effect of circSLC4A7 on DR was also confirmed by xenograft experiments. Collectively, circSLC4A7 in resistant-cells-derived exosomes promotes DCT resistance of PCa via miR-1205/MAPT axis, which may provide a new treatment strategy for DR of PCa.

摘要

前列腺癌 (PCa) 是一种高死亡率的癌症。多西他赛 (DCT) 联合第二代抗雄激素被认为是 PCa 的金标准治疗方法，但由于 DCT 耐药 (DR)，其应用受到限制。因此，探索 DR 的机制具有重要意义。在这项研究中，我们使用了 PC3 和 DU145 两种 PCa 细胞系，并通过用递增强度的 DCT 处理来构建 DR 细胞。建立了 circSLC4A7、miR-1205 和微管相关蛋白 tau (MAPT) 的转染。通过细胞计数试剂盒-8 测定法评估细胞活性和 DCT 的 IC。用 DCT 处理后，通过集落形成测定、流式细胞术分析和末端转移酶介导的 UTP 缺口末端标记测定来评估 DR。实时定量 PCR 和 Western blot 分析评估基因的表达水平。双荧光素酶报告基因测定验证了 circSLC4A7 与 MAPT 之间的 miR-1205 结合位点。进行了动物实验以评估 circSLC4A7 对肿瘤生长的影响。在构建 DR 细胞和分离外泌体后，我们发现不仅与 DR 细胞共培养，而且用 DR 细胞的外泌体处理也会促进正常细胞的 DR。此外，DR 细胞及其外泌体中高表达 circSLC4A7。circSLC4A7 的过表达增强了 DR，表现为 DCT 的 IC50 升高、DCT 处理后集落形成增加和细胞凋亡减少，而 circSLC4A7 的敲低则产生相反的效果。miR-1205 被证实是 circSLC4A7 的海绵 miRNA，miR-1205 抑制剂逆转了 sh-circSLC4A7 的作用。MAPT 进一步被确定为 miR-1205 的靶标，并且与 circSLC4A7 具有相似的作用。外泌体来源的 circSLC4A7 通过 miR-1205/MAPT 轴促进了 DR 细胞的 DCT 耐药性，这可能为 PCa 的 DR 提供了一种新的治疗策略。