背景:微RNA(miRNA),作为一种不可或缺的非编码RNA(ncRNA),参与不同的生物过程。然而,某些miRNA在胰腺导管腺癌(PDAC)中的具体调控机制尚不清楚.
方法:用RNA-scope和实时定量PCR(RT-qPCR)检测miR-194-5p在PDAC组织芯片和细胞系中的表达。通过几个功能实验观察了miR-194-5p在体外和体内携带的增殖和迁移功能。应用信息学方法和RNA测序数据探索miR-194-5p的靶标和miR-194-5p的上游环状RNA(circRNA)。RNA结合蛋白免疫沉淀(RIP)测定和双荧光素酶报告基因测定证实了miR-194-5p和SOCS2或miR-194-5p和circPVRL3之间的关系。通过救援实验获得了SOCS2和circPVRL3的增殖和迁移能力。
结果:在这项研究中,我们旨在阐明miR-194-5p的分子机制,在PDAC进展过程中具有关键作用。我们发现miR-194-5p在PDAC组织中的表达与癌旁组织相比显著上调,并且根据RNAscope和RT-qPCR与年龄和神经侵袭高度相关。miR-194-5p的过表达加速了细胞周期并增强了增殖和迁移过程。具体来说,circPVRL3,miR-194-5p,根据信息学方法,SOCS2被证实作为竞争性内源性RNA(ceRNA)工作,RIP,和双荧光素酶报告基因测定。此外,拯救实验证实了miR-194-5p之间的关系,circPVRL3和SOCS2mRNA。最后,circPVRL3/miR-194-5p/SOCS2轴激活PI3K/AKT信号通路调节PDAC的增殖和转移。
结论:我们的发现表明,circPVRL3下调引起的miR-194-5p增加刺激PI3K/AKT信号通路,通过circPVRL3/miR-194-5p/SOCS2轴促进PDAC进展,这表明circPVRL3/miR-194-5p/SOCS2轴可能是PDAC患者的潜在治疗靶点。
BACKGROUND: MicroRNAs (miRNAs), as an indispensable type of non-coding RNA (ncRNA), participate in diverse biological processes. However, the specific regulatory mechanism of certain miRNAs in pancreatic ductal adenocarcinoma (PDAC) remains unclear.
METHODS: The expression of miR-194-5p in PDAC tissue microarray and cell lines were detected by RNA-scope and real-time quantitative PCR (RT-qPCR). The function of proliferation and migration carried by miR-194-5p in vitro and vivo was observed by several functional experiments. Informatics methods and RNA sequencing data were applied to explore the target of miR-194-5p and the upstream circular RNA (circRNA) of miR-194-5p. RNA-binding protein immunoprecipitation (RIP) assay and dual-luciferase reporter assay confirmed the relationships between miR-194-5p and SOCS2 or miR-194-5p and circPVRL3. The proliferation and migration abilities of SOCS2 and circPVRL3 were accessed by rescue experiments.
RESULTS: In this study, we aimed to clarify the molecular mechanisms of miR-194-5p, which has critical roles during PDAC progression. We found that the expression of miR-194-5p was significantly upregulated in PDAC tissue compared to tumor-adjacent tissue and was highly related to age and nerve invasion according to RNAscope and RT‒qPCR. Overexpression of miR-194-5p accelerated the cell cycle and enhanced the proliferation and migration processes according to several functional experiments in vitro and in vivo. Specifically, circPVRL3, miR-194-5p, and SOCS2 were confirmed to work as competing endogenous RNAs (ceRNAs) according to informatics methods, RIP, and dual-luciferase reporter assays. Additionally, the rescue experiments confirmed the relationship among miR-194-5p, circPVRL3, and SOCS2 mRNA. Finally, the circPVRL3/miR-194-5p/SOCS2 axis activates the PI3K/AKT signaling pathway to regulate the proliferation and metastasis of PDAC.
CONCLUSIONS: Our findings indicated that an increase of miR-194-5p caused by circPVRL3 downregulation stimulates the PI3K/AKT signaling pathway to promote PDAC progression via the circPVRL3/miR-194-5p/SOCS2 axis, which suggests that the circPVRL3/miR-194-5p/SOCS2 axis may be a potential therapeutic target for PDAC patients.