Objective

This work aims to determine the role of circular RNA (circRNA) AGFG1 and related molecular mechanism in esophageal squamous cell carcinoma (ESCC) cells.

Methods

CircAGFG1 expression in ESCC cell lines was probed with qRT-PCR. ESCC cells were transfected/cotransfected with si-circAGFG1, pcDNA3.1-circAGFG1, si-Microtubule Associated Protein RP/EB Family Member 2 (MAPRE2), pcDNA3.1-circAGFG1 + miR-4306 mimic or pcDNA3.1-circAGFG1 + si-MAPRE2. The interactions between circAGFG1 and miR-4306 as well as miR-4306 and MAPRE2 were confirmed by dual-luciferase reporter assay. Cell proliferation, migration and invasion were detected by CCK-8, cell scratch and Transwell assays, respectively. Relative RNA expression levels of circAGFG1, miR-4306 and MAPRE2 in ESCC cells were measured by qRT-PCR. The protein level of MAPRE2 in ESCC cells was monitored by Western blot.

Results

CircAGFG1 was observably upregulated in ESCC cell lines. Besides, circAGFG1 silencing hindered ESCC cell development in vitro, and these effects were enhanced by miR-4306 overexpression or MAPRE2 silencing. Mechanistic analysis evidenced that circAGFG1 might act as a competitive endogenous RNA of miR-4306 to relieve the repressive effect of miR-4306 on its target MAPRE2.

Conclusion

CircAGFG1 facilitates ESCC progression via the miR-4306/MAPRE2 axis, and it may act as a possible biomarker for therapy and diagnosis in ESCC treatment.

中文翻译：

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CircAGFG1 充当 miR-4306 的海绵，通过调节 MAPRE2 表达来刺激食管癌进展

客观的

本工作旨在确定环状RNA（circRNA）AGFG1在食管鳞状细胞癌（ESCC）细胞中的作用及相关分子机制。

方法

用 qRT-PCR 探测 ESCC 细胞系中的 CircAGFG1 表达。用 si-circAGFG1、pcDNA3.1-circAGFG1、si-微管相关蛋白 RP/EB 家族成员 2 (MAPRE2)、pcDNA3.1-circAGFG1 + miR-4306 模拟物或 pcDNA3.1-circAGFG1 + si 转染/共转染 ESCC 细胞-MAPRE2。circAGFG1 和 miR-4306 以及 miR-4306 和 MAPRE2 之间的相互作用通过双荧光素酶报告基因分析得到证实。细胞增殖、迁移和侵袭分别通过CCK-8、细胞划痕和Transwell测定法检测。通过qRT-PCR测量ESCC细胞中circAGFG1、miR-4306和MAPRE2的相对RNA表达水平。通过蛋白质印迹监测ESCC细胞中MAPRE2的蛋白水平。

结果

CircAGFG1 在 ESCC 细胞系中明显上调。此外，circAGFG1 沉默阻碍了 ESCC 细胞的体外发育，而这些影响因 miR-4306 过表达或 MAPRE2 沉默而增强。机制分析表明，circAGFG1 可能作为 miR-4306 的竞争性内源性 RNA 来缓解 miR-4306 对其靶标 MAPRE2 的抑制作用。

结论

CircAGFG1 通过 miR-4306/MAPRE2 轴促进 ESCC 进展，它可以作为 ESCC 治疗中治疗和诊断的可能生物标志物。