Department of Hepatobiliary and Pancreatic Surgery, Hubei Cancer Hospital, Wuhan, China.
Department of Clinical Laboratory, Hubei Cancer Hospital, Wuhan, China.
Autoimmunity. 2021 Nov;54(7):471-482. doi: 10.1080/08916934.2021.1963958. Epub 2021 Aug 19.
Pancreatic cancer is a lethal malignancy in both sexes throughout the world. Circular RNAs (circRNAs) have been implicated in the development of pancreatic cancer by operating as competing endogenous RNAs (ceRNAs). Here, we explored circ_0099999-mediated ceRNA activity in regulating pancreatic tumorigenesis.
Ribonuclease R (RNase R) and subcellular localization assays were utilized to characterize circ_0099999. The levels of circ_0099999, microRNA (miR)-330-5p, and fascin actin-bundling protein 1 (FSCN1) were gauged by quantitative real-time PCR (qRT-PCR) and western blot. Cell proliferation, colony formation, apoptosis, migration, and invasion were evaluated by the Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, and transwell assays, respectively. The levels of glucose consumption and lactate production were determined using the assay kits. A direct relationship between miR-330-5p and circ_0099999 or FSCN1 was validated by dual-luciferase reporter assay. Tumour xenograft assays were used to analyse the role of circ_0099999 .
Circ_0099999 was highly up-regulated in pancreatic cancer tissues and cells. Knockdown of circ_0099999 impeded cell proliferation, migration, invasion, glycolysis, and promoted apoptosis , as well as diminished tumour growth . Circ_0099999 targeted miR-330-5p, and miR-330-5p was a downstream mediator of circ_0099999 function. FSCN1 was a direct and functional target of miR-330-5p. Furthermore, circ_0099999 operated as a ceRNA for miR-330-5p to modulate FSCN1 expression.
Our findings established a novel causal mechanism, circ_0099999/miR-330-5p/FSCN1 ceRNA crosstalk, in regulating pancreatic carcinogenesis and provided that inhibition of circ_0099999 might have therapeutic benefits in pancreatic cancer.
在全球范围内，胰腺癌在两性中都是一种致命的恶性肿瘤。环状 RNA（circRNA）已被证明通过作为竞争性内源性 RNA（ceRNA）参与胰腺癌的发展。在这里，我们探讨了 circ_0099999 介导的 ceRNA 活性在调节胰腺肿瘤发生中的作用。
利用核糖核酸酶 R（RNase R）和亚细胞定位测定来表征 circ_0099999。通过定量实时 PCR（qRT-PCR）和蛋白质印迹法测定 circ_0099999、微小 RNA（miR）-330-5p 和 fascin 肌动蛋白结合蛋白 1（FSCN1）的水平。通过细胞计数试剂盒-8（CCK-8）、集落形成、流式细胞术和 Transwell 测定分别评估细胞增殖、集落形成、细胞凋亡、迁移和侵袭。使用测定试剂盒测定葡萄糖消耗和乳酸产生的水平。通过双荧光素酶报告基因测定验证 miR-330-5p 与 circ_0099999 或 FSCN1 之间的直接关系。利用肿瘤异种移植实验分析 circ_0099999 的作用。
circ_0099999 在胰腺癌组织和细胞中高度上调。circ_0099999 的敲低抑制了细胞增殖、迁移、侵袭、糖酵解并促进了细胞凋亡，同时减少了肿瘤生长。circ_0099999 靶向 miR-330-5p，miR-330-5p 是 circ_0099999 功能的下游调节剂。FSCN1 是 miR-330-5p 的直接功能性靶标。此外，circ_0099999 作为 miR-330-5p 的 ceRNA 调节 FSCN1 的表达。
我们的研究结果确立了一种新的因果机制，即 circ_0099999/miR-330-5p/FSCN1 ceRNA 串扰，调节胰腺癌变，并表明抑制 circ_0099999 可能对胰腺癌具有治疗益处。
