Background/Objectives:Pancreatic cancer (PC) is among the most aggressive malignancies, often diagnosed at late stages. MicroRNAs (miRNAs) and proteins released from the tumor microenvironment into body fluids represent promising non-invasive biomarkers for early cancer detection. In this study, we took advantage of an innovative ultrasound (US)-based instrument (SonoWell®, Inno-Sol srl, Rome, Italy) to treat PC cells in order to promote and amplify the release of molecules, with the aim of identifying novel putative diagnostic PC biomarkers.Methods:Three human pancreatic adenocarcinoma cell lines (T3M-4, Panc02.03, and PaCa-44) and a non-cancerous pancreatic epithelial line (HPanEPic) were subjected to US using the SonoWell instrument. MiRNAs released in the supernatants were profiled by TaqMan-based qRT-PCR microfluidic cards, while proteins were analyzed by antibody arrays. Publicly available datasets of circulating miRNAs in PC patients were also reviewed.Results:Expression levels of 22 miRNAs in T3M-4 cells, 11 in Panc02.03, and 22 in PaCa-44, none of which were identified in the non-cancerous cell line profiling, were increased in the supernatant of US-treated as opposed to control cells. Among the statistically significant miRNAs or miRNAs common to at least two tumor cell lines, the expression levels of miR-155-5p, miR-320a, miR-32-5p, and miR-93-5p were also found to be significantly upregulated in sera from PC patients compared to the results for healthy controls. With regard to proteins released after sonication, several molecules were identified as candidate biomarkers in cancer US supernatants (Beta-2 microglobulin, CA125, CA19-9, CEA, CRP, Galectin-3, TIMP-1, uPA, and VEGF-A).Conclusions:We demonstrated that US-mediated sonoporation can promote and amplify the release of small molecules, miRNAs, and proteins into cell culture supernatants for consideration as putative biomarkers, thus encouraging further studies aimed at directly validating their expression levels in sera/plasma from PC patients and at deepening their role in the treatment of PC.
背景/目的：胰腺癌是最具侵袭性的恶性肿瘤之一，通常诊断时已处于晚期。从肿瘤微环境释放到体液中的微小RNA和蛋白质是极具前景的早期癌症检测非侵入性生物标志物。本研究利用创新型超声仪器（SonoWell®，意大利罗马Inno-Sol srl公司）处理胰腺癌细胞，以促进和放大分子释放，旨在鉴定新型潜在的胰腺癌诊断生物标志物。 方法：使用SonoWell仪器对三种人胰腺腺癌细胞系（T3M-4、Panc02.03和PaCa-44）及非癌性胰腺上皮细胞系（HPanEPic）进行超声处理。通过基于TaqMan的qRT-PCR微流控芯片分析上清液中释放的miRNA谱，同时采用抗体芯片分析蛋白质。此外，系统回顾了公开的胰腺癌患者循环miRNA数据集。 结果：超声处理组细胞上清液中，T3M-4细胞有22种miRNA、Panc02.03细胞有11种、PaCa-44细胞有22种表达水平升高，这些miRNA在非癌细胞系谱中均未检测到。在具有统计学显著性或至少两种肿瘤细胞系共有的miRNA中，miR-155-5p、miR-320a、miR-32-5p和miR-93-5p在胰腺癌患者血清中的表达水平较健康对照组显著上调。关于超声处理后释放的蛋白质，在癌症细胞超声上清液中鉴定出多个候选生物标志物分子（β2微球蛋白、CA125、CA19-9、CEA、CRP、半乳糖凝集素-3、TIMP-1、尿激酶型纤溶酶原激活物和血管内皮生长因子-A）。 结论：本研究证实超声介导的声孔效应能促进并放大细胞培养上清液中小分子、miRNA和蛋白质的释放，这些分子可作为潜在生物标志物。该发现鼓励进一步开展研究，直接验证其在胰腺癌患者血清/血浆中的表达水平，并深入探究其在胰腺癌治疗中的作用。
肿瘤(癌症)患者之家