Lee Kun-Lin, Liu Jun-Jen, Huang Wei-Jan, Hung Ching-Sheng, Liang Yu-Chih
Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, New Taipei City, Taiwan.
School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, 301 Yuantong Rd., New Taipei City, 23564, Taiwan.
Cancer Cell Int. 2025 Feb 19;25(1):56. doi: 10.1186/s12935-025-03687-0.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive types of solid tumor, and novel strategies must be developed for treating it. Previous studies predominantly utilized circular RNA (circRNA) expression plasmids incorporating Alu elements to facilitate the indirect expression of circRNA.
Public databases and bioinformatics tools were used to identify hsa_circ_0004781 that is highly expressed in PDAC and its potential microRNA (miRNA) targets and corresponding mRNA targets. Real hsa_circ_0004781, which is identical to the native form of hsa_circ_0004781 without any exogenous sequences, was prepared through in vitro transcription by using a ribozyme and ion-pair reversed-phase high-performance liquid chromatography (IP-RP HPLC). The biological functions of hsa_circ_0004781 were evaluated using loss-of-function and gain-of-function approaches with circRNA expression plasmids and real hsa_circ_0004781.
Knockdown of hsa_circ_0004781 inhibited the proliferation and migration of PDAC cells, whereas its overexpression produced opposite effects. Hsa_circ_0004781 was identified as a sponge for miR-9-5p and miR-338-3p, and its expression was negatively correlated with that of these miRNAs. Among the targets of miR-9-5p and miR-338-3p, Kruppel-like factor 5 (KLF5) and a disintegrin and metalloproteinase domain 17 (ADAM17) were negatively correlated with survival in patients with PDAC and were inversely regulated by these miRNAs. Furthermore, real hsa_circ_0004781 exhibited the same effects as those of the circRNA expression plasmids.
This study is the first to use real circRNAs to validate results obtained using circRNA expression plasmids. The results suggest that hsa_circ_0004781 functions as an oncogene, promoting the proliferation of PDAC cells through the miR-9-5p/KLF5 and miR-338-3p/ADAM17 axes. Therefore, hsa_circ_0004781 might be a therapeutic target for PDAC.
胰腺导管腺癌（PDAC）是侵袭性最强的实体瘤类型之一，必须开发新的治疗策略。以往的研究主要利用包含Alu元件的环状RNA（circRNA）表达质粒来促进circRNA的间接表达。
利用公共数据库和生物信息学工具鉴定在PDAC中高表达的hsa_circ_0004781及其潜在的微小RNA（miRNA）靶点和相应的信使核糖核酸（mRNA）靶点。通过使用核酶和离子对反相高效液相色谱（IP-RP HPLC）进行体外转录，制备了与天然形式的hsa_circ_0004781相同且无任何外源序列的真实hsa_circ_0004781。使用circRNA表达质粒和真实hsa_circ_0004781，通过功能缺失和功能获得方法评估hsa_circ_0004781的生物学功能。
敲低hsa_circ_0004781可抑制PDAC细胞的增殖和迁移，而其过表达则产生相反的效果。Hsa_circ_0004781被鉴定为miR-9-5p和miR-338-3p的海绵，其表达与这些miRNA的表达呈负相关。在miR-9-5p和miR-338-3p的靶点中， Kruppel样因子5（KLF5）和去整合素和金属蛋白酶结构域17（ADAM17）与PDAC患者的生存率呈负相关，并受到这些miRNA的反向调节。此外，真实的hsa_circ_0004781表现出与circRNA表达质粒相同的效果。
本研究首次使用真实的circRNAs验证使用circRNA表达质粒获得的结果。结果表明，hsa_circ_0004781作为一种癌基因，通过miR-9-5p/KLF5和miR-338-3p/ADAM17轴促进PDAC细胞的增殖。因此，hsa_circ_0004781可能是PDAC的治疗靶点。
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