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环状非编码 RNA circ_0072088 作为 ceRNA,靶向 miR-1225-5p/WT1 轴调控非小细胞肺癌细胞恶性行为。

Circular non-coding RNA circ_0072088 serves as a ceRNA, targeting the miR-1225-5p/WT1 axis to regulate non-small cell lung cancer cell malignant behavior.

机构信息

Emergency Department, Jingzhou Hospital Affiliated to Yangtze University, Jingzhou, China.

出版信息

Thorac Cancer. 2023 Jul;14(20):1969-1979. doi: 10.1111/1759-7714.14943. Epub 2023 May 23.

Abstract

BACKGROUND

Circular RNA (circRNA) circ_0072088 has been reported to be associated with NSCLC cell growth, migration, and invasion. However, the role and mechanism of circ_0072088 on NSCLC development have not yet been determined.

METHODS

Circ_0072088, microRNA-1225 (miR-1225-5p), and Wilms' tumor (WT1) suppressor gene level was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Migration, invasion, and apoptosis were detected using transwell and flow cytometry assays. Matrix metallopeptidase 9 (MMP9), hexokinase 2 (HK2), and WT1 were examined using western blot assay. The biological role of circ_0072088 on NSCLC tumor growth was examined by the xenograft tumor model in vivo. Circular RNA Interactome and TargetScan were used to predict the binding between miR-1225-5p and circ_0072088 or WT1, followed by confirmation using a dual-luciferase reporter.

RESULTS

Circ_0072088 and WT1 were highly expressed in NSCLC tissues and cells, and miR-1225-5p was decreased. Knockdown of circ_0072088 might repress migration, invasion, and glycolysis, and facilitate apoptosis of NSCLC cells in vitro. Circ_0072088 silencing also blocked NSCLC tumor growth in vivo. Mechanistically, circ_0072088 acted as a sponge of miR-1225-5p to regulate WT1 expression.

CONCLUSION

Circ_0072088 knockdown could inhibit cell growth, migration, invasion, and glycolysis partially by regulating the miR-1225-5p/WT1 axis, thus providing a promising therapeutic target for NSCLC treatment.

摘要

背景

环状 RNA(circRNA)circ_0072088 已被报道与非小细胞肺癌(NSCLC)细胞的生长、迁移和侵袭有关。然而,circ_0072088 对 NSCLC 发展的作用和机制尚未确定。

方法

采用逆转录定量聚合酶链反应(RT-qPCR)检测 circ_0072088、microRNA-1225(miR-1225-5p)和 Wilms 瘤(WT1)抑制基因水平。采用 Transwell 和流式细胞术检测迁移、侵袭和凋亡。采用 Western blot 检测基质金属蛋白酶 9(MMP9)、己糖激酶 2(HK2)和 WT1。采用体内异种移植肿瘤模型检测 circ_0072088 对 NSCLC 肿瘤生长的生物学作用。采用环状 RNA 相互作用组和 TargetScan 预测 miR-1225-5p 与 circ_0072088 或 WT1 的结合,随后采用双荧光素酶报告进行验证。

结果

circ_0072088 和 WT1 在 NSCLC 组织和细胞中高表达,miR-1225-5p 表达降低。circ_0072088 敲低可能抑制 NSCLC 细胞的迁移、侵袭和糖酵解,并促进其凋亡。circ_0072088 沉默也可抑制体内 NSCLC 肿瘤的生长。机制上,circ_0072088 作为 miR-1225-5p 的海绵调节 WT1 表达。

结论

circ_0072088 敲低通过调节 miR-1225-5p/WT1 轴可部分抑制细胞生长、迁移、侵袭和糖酵解,为 NSCLC 的治疗提供了有前景的治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dcf2/10344739/f1b71971817c/TCA-14-1969-g003.jpg

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