Department of Obstetrics and Gynecology, Faculty of Medicine, Medical Center‑University of Freiburg, D‑79106 Freiburg, Germany.
Institute of Medical Biometry and Statistics, Faculty of Medicine, Medical Center‑University of Freiburg, D‑79104 Freiburg, Germany.
Int J Oncol. 2020 Jan;56(1):47-68. doi: 10.3892/ijo.2019.4920. Epub 2019 Nov 25.
Due to the positive association between neoadjuvant chemotherapy (NACT) and the promising early response rates of patients with triple negative breast cancer (TNBC), including probabilities of pathological complete response, NACT is increasingly used in TNBC management. Liquid biopsy‑based biomarkers with the power to diagnose the early response to NACT may support established monitoring tools, which are to a certain extent imprecise and costly. Simple serum‑ or urine‑based analyses of non‑coding RNA (ncRNA) expression may allow for fast, minimally‑invasive testing and timely adjustment of the therapy regimen. The present study investigated breast cancer‑related ncRNAs [microRNA (miR)‑7, ‑9, ‑15a, ‑17, ‑18a, ‑19b, ‑21, ‑30b, ‑222 and ‑320c, PIWI‑interacting RNA‑36743 and GlyCCC2] in triple positive BT‑474 cells and three TNBC cell lines (BT‑20, HS‑578T and MDA‑MB‑231) treated with various chemotherapeutic agents using reverse transcription‑quantitative PCR. Intracellular and secreted microvesicular ncRNA expression levels were analysed using a multivariable statistical regression analysis. Chemotherapy‑driven effects were investigated by analysing cell cycle determinants at the mRNA and protein levels. Serum and urine specimens from 8 patients with TNBC were compared with 10 healthy females using two‑sample t‑tests. Samples from the patients with TNBC were compared at two time points. Chemotherapeutic treatments induced distinct changes in ncRNA expression in TNBC cell lines and the BT‑474 cell line in intra‑ and extracellular compartments. Serum and urine‑based ncRNA expression analysis was able to discriminate between patients with TNBC and controls. Time point comparisons in the urine samples of patients with TNBC revealed a general rise in the level of ncRNA. Serum data suggested a potential association between piR‑36743, miR‑17, ‑19b and ‑30b expression levels and an NACT‑driven complete clinical response. The present study highlighted the potential of ncRNAs as liquid biopsy‑based biomarkers in TNBC chemotherapy treatment. The ncRNAs tested in the present study have been previously investigated for their involvement in BC or TNBC chemotherapy responses; however, these previous studies were restricted to patient tissue or in vitro models. The data from the present study offer novel insight into ncRNA expression in liquid samples from patients with TNBC, and the study serves as an initial step in the evaluation of ncRNAs as diagnostic biomarkers in the monitoring of TNBC therapy.
由于新辅助化疗 (NACT) 与三阴性乳腺癌 (TNBC) 患者有希望的早期反应率之间存在正相关，包括病理完全缓解的概率，NACT 越来越多地用于 TNBC 管理。具有诊断 NACT 早期反应能力的基于液体活检的生物标志物可能支持现有的监测工具，这些监测工具在某种程度上不够精确且成本高昂。基于非编码 RNA (ncRNA) 表达的简单血清或尿液分析可能允许快速、微创检测和及时调整治疗方案。本研究使用逆转录定量 PCR 检测了三种 TNBC 细胞系 (BT-20、HS-578T 和 MDA-MB-231) 中与乳腺癌相关的 ncRNA [miR-7、-9、-15a、-17、-18a、-19b、-21、-30b、-222 和 -320c、PIWI 相互作用 RNA-36743 和 GlyCCC2]，以及用各种化疗药物处理的三阳性 BT-474 细胞。使用多变量统计回归分析分析了细胞内和分泌的微小囊泡 ncRNA 表达水平。通过分析 mRNA 和蛋白质水平的细胞周期决定因素来研究化疗驱动的作用。使用两样本 t 检验将 8 名 TNBC 患者的血清和尿液标本与 10 名健康女性进行比较。使用两样本 t 检验比较了 TNBC 患者的两个时间点的标本。化疗在 TNBC 细胞系和 BT-474 细胞系的细胞内和细胞外区室中诱导了 ncRNA 表达的明显变化。基于血清和尿液的 ncRNA 表达分析能够区分 TNBC 患者和对照组。TNBC 患者尿液样本的时间点比较显示 ncRNA 水平普遍升高。血清数据表明 piR-36743、miR-17、-19b 和 -30b 表达水平与 NACT 驱动的完全临床反应之间可能存在关联。本研究强调了 ncRNA 作为 TNBC 化疗治疗中基于液体活检的生物标志物的潜力。本研究中测试的 ncRNA 先前已被研究用于其在 BC 或 TNBC 化疗反应中的作用；然而，这些先前的研究仅限于患者组织或体外模型。本研究的数据提供了关于 TNBC 患者液体样本中 ncRNA 表达的新见解，并作为评估 ncRNA 作为 TNBC 治疗监测的诊断生物标志物的初步步骤。
