长链非编码 RNA TDRKH-AS1 通过 miR-134-5p/CREB1 轴促进乳腺癌进展。
LncRNA TDRKH-AS1 promotes breast cancer progression via the miR-134-5p/CREB1 axis.
机构信息
Department of Breast Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, Zhejiang, China.
Department of Breast Surgery, Zhejiang Cancer Hospital, Institute of Cancer and Basic Medicine (IBMC), Chinese Academy of Sciences, Hangzhou, 310022, Zhejiang, China.
出版信息
J Transl Med. 2023 Nov 26;21(1):854. doi: 10.1186/s12967-023-04640-3.
BACKGROUND
Breast cancer (BC) is a prevalent malignancy with complex etiology and varied clinical behavior. Long non-coding RNAs (lncRNAs) have emerged as key regulators in cancer progression, including BC. Among these, lncRNA TDRKH-AS1 has been implicated in several cancers, but its role in BC remains unclear.
METHODS
We conducted a comprehensive investigation to elucidate the role of TDRKH-AS1 in BC. Clinical samples were collected from BC patients, and BC cell lines were cultured. Bioinformatics analysis using the starBase database was carried out to assess TDRKH-AS1 expression levels in BC tissue samples. Functional experiments, including knockdown, colony formation, CCK-8, Transwell, and wound-healing assays, were conducted to determine the role of TDRKH-AS1 in BC cell proliferation and invasion. Luciferase reporter and RIP assays were used to examine the interactions between TDRKH-AS1 and miR-134-5p. In addition, the downstream target gene of miR-134-5p, cAMP response element-binding protein 1 (CREB1), was identified and studied using various methods, including RT-qPCR, immunoprecipitation, and rescue experiments. In vivo experiments using mouse tumor xenograft models were conducted to examine the role of TDRKH-AS1 in BC tumorigenesis.
RESULTS
TDRKH-AS1 was found to be significantly upregulated in BC tissues and cell lines. High TDRKH-AS1 expression correlated with advanced BC stages and worse patient outcomes. Knockdown of TDRKH-AS1 led to decreased BC cell proliferation and invasion. Mechanistically, TDRKH-AS1 acted as a sponge for miR-134-5p, thereby reducing the inhibitory effects of miR-134-5p on CREB1 expression. Overexpression of CREB1 partially rescued the effects of TDRKH-AS1 knockdown in BC cells. In vivo studies further confirmed the tumor-promoting role of TDRKH-AS1 in BC.
CONCLUSIONS
Our study unveiled a novel regulatory axis involving TDRKH-AS1, miR-134-5p, and CREB1 in BC progression. TDRKH-AS1 functioned as an oncogenic lncRNA by promoting BC cell proliferation and invasion through modulation of the miR-134-5p/CREB1 axis. These findings highlighted TDRKH-AS1 as a potential diagnostic biomarker and therapeutic target for BC treatment.
背景
乳腺癌（BC）是一种常见的恶性肿瘤，其病因复杂，临床表现多样。长链非编码 RNA（lncRNAs）已成为癌症进展的关键调节因子，包括 BC。在这些 lncRNAs 中，lncRNA TDRKH-AS1 已被发现在多种癌症中发挥作用，但在 BC 中的作用尚不清楚。
方法
我们进行了一项全面的研究，以阐明 TDRKH-AS1 在 BC 中的作用。从 BC 患者中收集临床样本，并培养 BC 细胞系。使用 starBase 数据库进行生物信息学分析，评估 TDRKH-AS1 在 BC 组织样本中的表达水平。进行功能实验，包括敲低、集落形成、CCK-8、Transwell 和划痕愈合实验，以确定 TDRKH-AS1 在 BC 细胞增殖和侵袭中的作用。使用荧光素酶报告和 RIP 实验来检测 TDRKH-AS1 与 miR-134-5p 之间的相互作用。此外，使用各种方法，包括 RT-qPCR、免疫沉淀和拯救实验，鉴定和研究 miR-134-5p 的下游靶基因 cAMP 反应元件结合蛋白 1（CREB1）。使用小鼠肿瘤异种移植模型进行体内实验，以研究 TDRKH-AS1 在 BC 肿瘤发生中的作用。
结果
发现 TDRKH-AS1 在 BC 组织和细胞系中显著上调。高 TDRKH-AS1 表达与晚期 BC 分期和患者预后不良相关。敲低 TDRKH-AS1 导致 BC 细胞增殖和侵袭减少。机制上，TDRKH-AS1 作为 miR-134-5p 的海绵，从而降低 miR-134-5p 对 CREB1 表达的抑制作用。CREB1 的过表达部分挽救了 TDRKH-AS1 敲低对 BC 细胞的作用。体内研究进一步证实了 TDRKH-AS1 在 BC 中的促肿瘤作用。
结论
我们的研究揭示了一个涉及 TDRKH-AS1、miR-134-5p 和 CREB1 的新型调控轴在 BC 进展中的作用。TDRKH-AS1 通过调节 miR-134-5p/CREB1 轴促进 BC 细胞增殖和侵袭，发挥致癌 lncRNA 的作用。这些发现凸显了 TDRKH-AS1 作为 BC 治疗的潜在诊断生物标志物和治疗靶点的潜力。
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