A Novel circ_0075829/miR-326/GOT1 ceRNA Crosstalk Regulates the Malignant Phenotypes and Drug Sensitivity of Gemcitabine-Resistant Pancreatic Cancer Cells

IF 3.2 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yongjia Xiang, Rubing Zhou, Yi Yang, Hao Bai, Fan Liang, Hongmei Wang, Xia Wang
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Abstract

Although gemcitabine (GEM) is the cornerstone of the treatment of pancreatic cancer (PC), GEM resistance frequently arises. Circular RNA (circRNA) circ_0075829 is highly expressed in PC. However, whether circ_0075829 contributes to GEM resistance of PC is largely unknown. To generate GEM-resistant PC cells (BxPC-3/GR and SW1990/GR), we exposed GEM-sensitive PC cells to GEM. Circ_0075829, microRNA (miR)-326, and glutamic-oxaloacetic transaminase 1 (GOT1) were quantified by a qRT-PCR or western blot method. Cell survival and viability were gauged by MTS assay. Cell proliferation, apoptosis, invasion, and migration were assessed by EdU, flow cytometry, transwell, and wound-healing assays, respectively. Dual-luciferase reporter assays were used to verify the relationship between miR-326 and circ_0075829 or GOT1. Mouse xenografts were performed to evaluate the role of circ_0075829 in vivo. Our data showed that circ_0075829 was upregulated in GEM-resistant PC tissues and cells. Knockdown of circ_0075829 impeded the proliferation, invasion, migration, and glutamine metabolism, and promoted cell apoptosis and GEM sensitivity of GEM-resistant PC cells. Moreover, circ_0075829 silencing suppressed the tumorigenicity of SW1990/GR cells and sensitized them to the cytotoxic effect of GME in vivo. Mechanistically, circ_0075829 bound miR-326 and exerted regulatory effects by affecting miR-326 expression. GOT1 was a direct miR-326 target and a key downstream effector of miR-326. Furthermore, circ_0075829 modulated GOT1 expression via miR-326. Our findings establish a novel regulatory network, the circ_0075829/miR-326/GOT1 competing endogenous RNA (ceRNA) crosstalk, in the regulation of GEM resistance in PC.

Abstract Image

尽管吉西他滨(GEM)是治疗胰腺癌(PC)的基石,但经常会出现 GEM 抗药性。环状 RNA(circRNA)circ_0075829 在 PC 中高度表达。然而,circ_0075829是否会导致PC对GEM产生耐药性,目前尚不清楚。为了生成抗 GEM 的 PC 细胞(BxPC-3/GR 和 SW1990/GR),我们将对 GEM 敏感的 PC 细胞暴露于 GEM。我们采用 qRT-PCR 或 Western 印迹法对 Circ_0075829、microRNA (miR)-326 和谷氨酸-草酰乙酸转氨酶 1 (GOT1) 进行了定量分析。细胞存活率和活力通过 MTS 法检测。细胞增殖、凋亡、侵袭和迁移分别通过 EdU、流式细胞仪、transwell 和伤口愈合试验进行评估。双荧光素酶报告实验用于验证 miR-326 与 circ_0075829 或 GOT1 之间的关系。小鼠异种移植实验评估了 circ_0075829 在体内的作用。我们的数据显示,circ_0075829 在耐 GEM 的 PC 组织和细胞中上调。敲除circ_0075829会阻碍GEM耐药PC细胞的增殖、侵袭、迁移和谷氨酰胺代谢,促进细胞凋亡和对GEM的敏感性。此外,沉默circ_0075829还抑制了SW1990/GR细胞的致瘤性,并使其对体内GME的细胞毒性作用敏感。从机制上讲,circ_0075829与miR-326结合,并通过影响miR-326的表达发挥调控作用。GOT1 是 miR-326 的直接靶标,也是 miR-326 的关键下游效应物。此外,circ_0075829 通过 miR-326 调节 GOT1 的表达。我们的研究结果建立了一个新的调控网络,即circ_0075829/miR-326/GOT1竞争性内源性RNA(ceRNA)串联调控PC的GEM抗性。
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来源期刊
CiteScore
5.80
自引率
2.80%
发文量
277
审稿时长
6-12 weeks
期刊介绍: The Journal of Biochemical and Molecular Toxicology is an international journal that contains original research papers, rapid communications, mini-reviews, and book reviews, all focusing on the molecular mechanisms of action and detoxication of exogenous and endogenous chemicals and toxic agents. The scope includes effects on the organism at all stages of development, on organ systems, tissues, and cells as well as on enzymes, receptors, hormones, and genes. The biochemical and molecular aspects of uptake, transport, storage, excretion, lactivation and detoxication of drugs, agricultural, industrial and environmental chemicals, natural products and food additives are all subjects suitable for publication. Of particular interest are aspects of molecular biology related to biochemical toxicology. These include studies of the expression of genes related to detoxication and activation enzymes, toxicants with modes of action involving effects on nucleic acids, gene expression and protein synthesis, and the toxicity of products derived from biotechnology.
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