环状 RNA circ_0006089 通过调控 miR-143-3p/PTBP3 轴和 PI3K/AKT 信号通路促进胃癌的进展。

Circular RNA circ_0006089 promotes the progression of gastric cancer by regulating the miR-143-3p/PTBP3 axis and PI3K/AKT signaling pathway.

机构信息

Department of General Surgery, The Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province, China.

出版信息

J Dig Dis. 2022 Jul;23(7):376-387. doi: 10.1111/1751-2980.13116. Epub 2022 Aug 23.

DOI:10.1111/1751-2980.13116

PMID:35844201

Abstract

OBJECTIVES

Circular RNAs (circRNAs) play pivotal roles in malignancies including gastric cancer (GC). We aimed to investigate the biological function and regulatory mechanism of circ_0006089 in GC.

METHODS

Circ_0006089, microRNA (miR)-143-3p, and polypyrimidine tract-binding protein 3 (PTBP3) expressions were measured via quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in GC cell lines. Cell proliferative capacity was determined by colony formation and CCK-8 assays. Flow cytometry was employed for measuring cell apoptosis. Cell invasion and migration were measured via transwell and wound-healing assays. Western blot analysis was utilized for detecting protein expressions of E-cadherin, N-cadherin, vimentin, PTBP3, PI3K, p-PI3K, AKT, and p-AKT. Dual-reporter luciferase analysis was conducted to confirm the association between miR-143-3p and circ_0006089 or PTBP3. The role of circ_0006089 in vivo was detected via establishing a mice xenograft model.

RESULTS

Circ_0006089 expression was increased in GC. Circ_0006089 downregulation suppressed the proliferation and metastasis and induced apoptosis of GC cells, which was counteracted by miR-143-3p inhibition or PTBP3 overexpression. In addition, circ_0006089 overexpression could promote GC progression. MiR-143-3p specially bound to circ_0006089 and PTBP3 was targeted by miR-143-3p. Moreover, circ_0006089 could regulate PTBP3 expression and the PI3K/AKT pathway by sponging miR-143-3p. Circ_0006089 knockdown also suppressed tumor growth.

CONCLUSION

Circ_0006089 regulated miR-143-3p/PTBP3/PI3K/AKT pathway to facilitate GC progression.

摘要

目的

环状 RNA（circRNAs）在包括胃癌（GC）在内的多种恶性肿瘤中发挥着关键作用。本研究旨在探究 circ_0006089 在 GC 中的生物学功能和调控机制。

方法

采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 GC 细胞系中 circ_0006089、微小 RNA（miR）-143-3p 和多嘧啶 tract 结合蛋白 3（PTBP3）的表达。通过集落形成和 CCK-8 检测评估细胞增殖能力。采用流式细胞术检测细胞凋亡。通过 Transwell 和划痕愈合实验检测细胞侵袭和迁移能力。Western blot 分析用于检测 E-钙黏蛋白、N-钙黏蛋白、波形蛋白、PTBP3、PI3K、p-PI3K、AKT 和 p-AKT 的蛋白表达。双荧光素酶报告基因分析用于验证 miR-143-3p 与 circ_0006089 或 PTBP3 的关联。通过建立小鼠异种移植模型检测 circ_0006089 体内的作用。

结果

circ_0006089 在 GC 中表达上调。circ_0006089 下调抑制 GC 细胞的增殖和转移，并诱导其凋亡，该作用可被 miR-143-3p 抑制或 PTBP3 过表达所拮抗。此外，circ_0006089 过表达可促进 GC 进展。miR-143-3p 特异性结合 circ_0006089，PTBP3 是 miR-143-3p 的靶基因。此外，circ_0006089 通过海绵吸附 miR-143-3p 来调节 PTBP3 表达和 PI3K/AKT 通路。circ_0006089 下调也抑制了肿瘤生长。