通过血清胞外囊泡包裹的长非编码 RNA HEVEPA 进行胰腺癌液体活检。

IF 1.7 4区 医学 Q3 GASTROENTEROLOGY & HEPATOLOGY Pancreas Pub Date : 2024-05-01 Epub Date: 2024-02-26 DOI:10.1097/MPA.0000000000002315
Kenji Takahashi, Tatsutoshi Inuzuka, Yuta Shimizu, Kazuki Sawamoto, Kenzui Taniue, Yusuke Ono, Fumi Asai, Kazuya Koyama, Hiroki Sato, Hidemasa Kawabata, Hidetaka Iwamoto, Keisuke Yamakita, Yohei Kitano, Takashi Teramoto, Mikihiro Fujiya, Satoshi Fujii, Yusuke Mizukami, Toshikatsu Okumura
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引用次数: 0

摘要

研究目的长非编码 RNA(lncRNA)在胰腺导管腺癌(PDAC)中的作用仍不清楚。细胞外囊泡 (EV) 包被的 RNA 可作为液体活检的有效靶标。我们旨在鉴定PDAC中以前未知的EV包被的lncRNA,并建立高精度的EV分离方法:使用现有的和新开发的方法,即 PEViA-UC 和 PEViA-IP,从 20 例 PDAC 患者、22 例导管内乳头状黏液瘤患者和 21 例健康人的血清样本中分离胞外囊泡。使用数字 PCR 分析了细胞外囊泡 lncRNA 的表达:结果:使用cDNA芯片进行的基因表达分析表明,PDAC患者血清EVs中存在一种高表达的lncRNA--HEVEPA。我们利用 PEViA 试剂、超速离心和免疫沉淀技术建立了 PEViA-UC 和 PEViA-IP。尽管使用现有方法检测EV包裹的HEVEPA具有挑战性,但PEViA-UC和PEViA-IP检测到了EV HEVEPA,与非PDAC患者相比,EV HEVEPA在PDAC患者中高表达。与单独检测 HEVEPA、HULC 或 CA19-9 相比,使用 HEVEPA 和 HULC(PDAC 中高表达的 lncRNA)以及碳水化合物抗原 19-9(CA19-9)组合检测 PDAC 和非 PDAC 的检测灵敏度更高:结论:利用 PEViA-IP 和 CA19-9 分离出的细胞外囊泡 lncRNA 可成为液体活检诊断 PDAC 的有效靶标。
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Liquid Biopsy for Pancreatic Cancer by Serum Extracellular Vesicle-Encapsulated Long Noncoding RNA HEVEPA.

Objectives: The role of long noncoding RNAs (lncRNAs) in pancreatic ductal adenocarcinoma (PDAC) remain unclear. Extracellular vesicle (EV)-encapsulated RNAs could be effective targets for liquid biopsy. We aimed to identify previously unknown EV-encapsulated lncRNAs in PDAC and establish highly accurate methods for isolating EVs.

Materials and methods: Extracellular vesicles were isolated using existing and newly developed methods, namely, PEViA-UC and PEViA-IP, from serum samples of 20 patients with PDAC, 22 patients with intraductal papillary mucinous neoplasms, and 21 healthy individuals. Extracellular vesicle lncRNA expression was analyzed using digital PCR.

Results: Gene expression analysis using cDNA microarray revealed a highly expressed lncRNA, HEVEPA , in serum EVs from patients with PDAC. We established PEViA-UC and PEViA-IP using PEViA reagent, ultracentrifugation, and immunoprecipitation. Although detection of EV-encapsulated HEVEPA using existing methods is challenging, PEViA-UC and PEViA-IP detected EV HEVEPA , which was highly expressed in patients with PDAC compared with non-PDAC patients. The detection sensitivity for discriminating PDAC from non-PDAC using the combination of HEVEPA and HULC , which are highly expressed lncRNAs in PDAC, and carbohydrate antigen 19-9 (CA19-9), was higher than that of HEVEPA , HULC , or CA19-9 alone.

Conclusions: Extracellular vesicle lncRNAs isolated using PEViA-IP and CA19-9 together could be effective targets in liquid biopsy for PDAC diagnosis.

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来源期刊
Pancreas
Pancreas 医学-胃肠肝病学
CiteScore
4.70
自引率
3.40%
发文量
289
审稿时长
1 months
期刊介绍: Pancreas provides a central forum for communication of original works involving both basic and clinical research on the exocrine and endocrine pancreas and their interrelationships and consequences in disease states. This multidisciplinary, international journal covers the whole spectrum of basic sciences, etiology, prevention, pathophysiology, diagnosis, and surgical and medical management of pancreatic diseases, including cancer.
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