基于全转录组测序分析的探索三阴性乳腺癌发病机制和治疗靶点的淋巴转移相关环状 RNA—miRNA—mRNA 网络：一项实验验证研究。
Lymphatic metastasis-associated circRNA‒miRNA‒mRNA network for exploring the pathogenesis and therapeutic target of triple negative breast cancer based on whole-transcriptome sequencing analysis: an experimental verification study.
机构信息
Department of Breast Surgery, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, 510623, Guangdong, China.
Department of Anesthesiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120, Guangdong, China.
出版信息
J Transl Med. 2022 Nov 5;20(1):508. doi: 10.1186/s12967-022-03728-6.
BACKGROUND
The metastatic mechanisms of axillary lymph nodes (ALNs) in triple-negative breast cancer (TNBC) remain unclear. We aimed to identify the potential circRNA regulatory network in ALN metastasis.
METHODS
We performed whole transcriptome sequencing (WTS) to determine the expression profiles of RNAs and screen out differentially expressed messenger RNAs (DEMs), microRNAs (DEMis), and circRNAs (DECs) between ALN-positive and ALN-negative TNBC patients. Functional enrichment analysis and Kaplan-Meier survival analysis were utilized to unearth the potential regulatory mechanisms of the DEMs. A competing endogenous RNA (ceRNA) network was constructed using computational biology. The expression levels of DECs in cell lines were confirmed by real-time polymerase chain reaction (RT‒PCR).
RESULTS
Following WTS and differential expression analysis, 739 DEMs, 110 DEMis, and 206 DECs were identified between ALN-positive and ALN-negative TNBC patients. Functional analysis indicated that the DEMs mainly functioned in carcinogenesis and tumor progression-related pathways. ceRNA networks containing eight circRNAs, six miRNAs, and eighteen mRNAs were developed. In the ceRNA network, two mRNAs (RAB3D and EDARADD) that were significantly associated with better overall survival and one mRNA (GSR) that predicted favorable recurrence-free survival in TNBC patients were chosen for further analysis. Then, a survival-related ceRNA network containing two DECs (hsa_circ_0061260 and hsa_circ_0060876), two DEMis (hsa-miR-5000-3p and hsa-miR-4792), and three mRNAs (GSR, RAB3D, and EDARADD) was identified. Then, two candidate DECs were validated by real-time PCR.
CONCLUSION
Our research constructed a ceRNA network that provides novel insights into the molecular mechanism of ALN metastasis and potential therapeutic targets in TNBC.
背景
三阴性乳腺癌（TNBC）腋窝淋巴结（ALN）转移的机制仍不清楚。我们旨在确定 ALN 转移中潜在的 circRNA 调控网络。
方法
我们进行了全转录组测序（WTS），以确定 RNA 的表达谱，并筛选出 ALN 阳性和 ALN 阴性 TNBC 患者之间差异表达的信使 RNA（DEM）、microRNA（DEM）和 circRNA（DEC）。功能富集分析和 Kaplan-Meier 生存分析用于揭示 DEM 的潜在调控机制。使用计算生物学构建竞争性内源性 RNA（ceRNA）网络。通过实时聚合酶链反应（RT-PCR）确认细胞系中 DEC 的表达水平。
结果
经过 WTS 和差异表达分析，在 ALN 阳性和 ALN 阴性 TNBC 患者之间鉴定出 739 个 DEM、110 个 DEM 和 206 个 DEC。功能分析表明，DEM 主要在致癌和肿瘤进展相关途径中发挥作用。构建了包含八个 circRNA、六个 miRNA 和十八个 mRNA 的 ceRNA 网络。在 ceRNA 网络中，两个与 TNBC 患者总生存期显著相关的 mRNA（RAB3D 和 EDARADD）和一个预测无复发生存率较好的 mRNA（GSR）被选择用于进一步分析。然后，选择了一个包含两个 DEC（hsa_circ_0061260 和 hsa_circ_0060876）、两个 DEM（hsa-miR-5000-3p 和 hsa-miR-4792）和三个 mRNAs（GSR、RAB3D 和 EDARADD）的与生存相关的 ceRNA 网络。然后，通过实时 PCR 验证了两个候选 DEC。
结论
我们的研究构建了 ceRNA 网络，为 ALN 转移的分子机制和 TNBC 的潜在治疗靶点提供了新的见解。
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