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膀胱癌进展受到心脏和神经嵴衍生物表达 2 反义 RNA 1/miRNA-93-5p/缺陷型 Cullin Neddylation 1 结构域包含 3 轴的抑制。

Bladder Cancer Progression Is Suppressed Through the Heart and Neural Crest Derivatives Expressed 2-Antisense RNA 1/microRNA-93-5p/Defective in Cullin Neddylation 1 Domain Containing 3 Axis.

机构信息

Provincial Clinical Medical College of Fujian Medical University, Fuzhou, 350001, China.

Department of Urology, Fujian Provincial Hospital, No. 516 Jinrong South Road, Cangshan District, Fuzhou, 350001, China.

出版信息

Appl Biochem Biotechnol. 2023 Jul;195(7):4116-4133. doi: 10.1007/s12010-022-04295-8. Epub 2023 Jan 19.

Abstract

MicroRNAs (miRNAs) are critical in progression of bladder cancer (BCa). miRNA-93-5p is increased in cancers and is positively correlated with an unfavorable prognosis. But its effects on BCa remain rarely understood. This investigation aimed to dig out miRNA-93-5p affecting biological behaviors of BCa. In this research, mRNA and protein expression in cancer cells were assessed via quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell Counting Kit-8 (CCK-8), colony formation, scratch healing, and transwell assays were utilized to analyze cancer cell viability, colony-forming, migration, and invasion, respectively. Bioinformatics analysis predicted upstream regulatory genes and downstream target genes of miRNA-93-5p, with the targeting relationship being verified through a dual-luciferase assay. The BCa xenograft model in nude mice further investigated the effect of miRNA-93-5p and AND2-AS1 on tumor size and quality, and validated the relationship between HAND2-AS1/miRNA-93-5p/DCUN1D3. Our results displayed that miRNA-93-5p was increased in BCa cell lines. Knockdown miRNA-93-5p constrained BCa cell malignant phenotypes. HAND2-AS1 targeted miRNA-93-5p, thus restraining malignant progression of BCa cells. DCUN1D3 was found downstream of miRNA-93-5p. miRNA-93-5p modulated proliferation, migration, and invasion of BCa cells by targeting DCUN1D3. In vivo experiments disclosed that forced expression of lncRNA HAND2-AS1, and inhibited miRNA-93-5p regressed tumor growth. Meanwhile, the same as the results of cell experiments, the expression of miRNA-93-5p was downregulated, and DCUN1D3 expression was advanced in tumor tissues. To conclude, lncRNA HAND2-AS1 exerted anti-tumor effects and regulated BCa cell proliferation, invasion, and migration by targeting miRNA-93-5p/DCUN1D3.

摘要

微小 RNA(miRNAs)在膀胱癌(BCa)的进展中起着关键作用。miRNA-93-5p 在癌症中增加,与不良预后呈正相关。但其对 BCa 的影响仍知之甚少。本研究旨在挖掘影响 BCa 生物学行为的 miRNA-93-5p。在这项研究中,通过定量实时聚合酶链反应(qRT-PCR)和 Western blot 评估癌细胞中的 mRNA 和蛋白质表达。细胞计数试剂盒-8(CCK-8)、集落形成、划痕愈合和 Transwell 测定分别用于分析癌细胞活力、集落形成、迁移和侵袭。生物信息学分析预测了 miRNA-93-5p 的上游调节基因和下游靶基因,通过双荧光素酶报告基因实验验证了靶向关系。裸鼠 BCa 移植瘤模型进一步研究了 miRNA-93-5p 和 AND2-AS1 对肿瘤大小和质量的影响,并验证了 HAND2-AS1/miRNA-93-5p/DCUN1D3 之间的关系。研究结果显示,miRNA-93-5p 在 BCa 细胞系中增加。敲低 miRNA-93-5p 抑制了 BCa 细胞的恶性表型。HAND2-AS1 靶向 miRNA-93-5p,从而抑制 BCa 细胞的恶性进展。DCUN1D3 是 miRNA-93-5p 的下游靶基因。miRNA-93-5p 通过靶向 DCUN1D3 调节 BCa 细胞的增殖、迁移和侵袭。体内实验表明,过表达 lncRNA HAND2-AS1 和抑制 miRNA-93-5p 均可抑制肿瘤生长。同时,与细胞实验结果一致,肿瘤组织中 miRNA-93-5p 表达下调,DCUN1D3 表达上调。综上所述,lncRNA HAND2-AS1 通过靶向 miRNA-93-5p/DCUN1D3 发挥抗肿瘤作用,并调节 BCa 细胞的增殖、侵袭和迁移。

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