Department of General Surgery, Wuhan Third Hospital (Tongren Hospital of Wuhan University).
Department of Oncology, Wuhan Asia General Hospital, Wuhan, China.
Anticancer Drugs. 2023 Oct 1;34(9):1002-1009. doi: 10.1097/CAD.0000000000001487. Epub 2023 Jan 24.
Enhancing research indicatedthat circular RNA (circRNA) acted a critical part in cholangiocarcinoma (CHOL) development. This research aims to discover the role of circRNA SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 5 (SMARCA5) in CHOL bio-progression, which has been proved to be downregulated in CHOL tissues. In this study, quantitative reverse transcription polymerase chain reaction was used to reveal the level and linkage of circRNA SMARCA5, miRNA-95-3p and TNF receptor-associated factor 3 gene (TRAF3) in CHOL tissues and cancer cells. The target sites of circRNA SMARCA5 and miRNA-95-3p were forecast by Starbase, and Targetscan was conducted to forecast the potential linkage points of TRAF3 and miRNA-95-3p, and which is affirmed by double luciferase reporter assay. CCK-8 and flow cytometry assay was carried to indicate cell viability. And apoptosis-related protein was counted by caspase3 activity and Western blot assay. CircRNA SMARCA5 was downregulated in CHOL cell lines and cancer samples. Besides, over-expression of SMARCA5 inhibited cell growth and promoted apoptotic rate. Dual-luciferase reporter assays presented that miRNA-95-3p could link with circRNA SMARCA5. Moreover, miRNA-95-3p was discovered highly expressed in CHOL. Interference of miRNA-95-3p repressed cell proliferation and raised the apoptosis. Importantly, TRAF3 was validated to be a downstream of miRNA-95-3p. Strengthen of miRNA-95-3p reversed the inhibitory impact of circRNA SMARCA5-plasmid transfection, and the results of miRNA-95-3p inhibitor were reversed by si-TRAF3. CircRNA SMARCA5 is involved in CHOL development by interosculating miRNA-95-3p/TRAF3 axis and may become a novel approach for treating CHOL.
研究表明环状 RNA（circRNA）在胆管癌（CHOL）发展中起着关键作用。本研究旨在发现 SWI/SNF 相关、基质相关、肌动蛋白依赖性染色质调节因子亚家族 A 成员 5（SMARCA5）circRNA 在 CHOL 生物进展中的作用，该基因已被证明在 CHOL 组织中下调。在这项研究中，使用定量逆转录聚合酶链反应揭示了 CHOL 组织和癌细胞中 circRNA SMARCA5、miRNA-95-3p 和肿瘤坏死因子受体相关因子 3 基因（TRAF3）的水平和关联。通过 Starbase 预测 circRNA SMARCA5 和 miRNA-95-3p 的靶位点，通过 Targetscan 预测 TRAF3 和 miRNA-95-3p 的潜在关联点，并通过双荧光素酶报告基因实验证实。CCK-8 和流式细胞术检测细胞活力。通过 caspase3 活性和 Western blot 检测计数凋亡相关蛋白。SMARCA5 circRNA 在 CHOL 细胞系和癌症样本中下调。此外，SMARCA5 的过表达抑制细胞生长并促进凋亡率。双荧光素酶报告基因实验表明，miRNA-95-3p 可以与 circRNA SMARCA5 结合。此外，在 CHOL 中发现 miRNA-95-3p 高表达。miRNA-95-3p 的干扰抑制细胞增殖并提高凋亡率。重要的是，TRAF3 被验证为 miRNA-95-3p 的下游靶基因。miRNA-95-3p 增强子逆转了 circRNA SMARCA5 质粒转染的抑制作用，而 miRNA-95-3p 抑制剂的结果则被 si-TRAF3 逆转。circRNA SMARCA5 通过相互作用 miRNA-95-3p/TRAF3 轴参与 CHOL 的发展，可能成为治疗 CHOL 的新方法。
