Department of Pancreatic and Endocrine Surgery, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(17):8845-8854. doi: 10.26355/eurrev_202009_22824.
The aim of this study is to investigate the expression levels of circRNA_100782 in gastric cancer tissues, and its function of regulating tumor suppressor gene Rb by absorbing miR-574-3p in a sponge form.
qRT-PCR was performed to detect the expressions of circRNA_100782 at different stages during gastric cancer tissues. CCK-8 assay was performed to evaluate the osteoclast proliferation and differentiation. The correlation between miR-574-3p and circRNA_100782 was detected by statistical analysis. Bioinformatics and Luciferase assay were performed to explore the interaction and binding site of circRNA_100782 and miR-574-3p. The mice Rb 3'-UTR were cloned into the Luciferase reporter vector and miR-574-3p binding mutants were constructed to validate the inhibited regulation of miR-574-3p to the expression of Rb.
In the current study, compared with adjacent non-cancerous normal tissues, the expressions of circRNA_100782 and Rb were both downregulated in human gastric cancer cells. Through qRT-PCR and CCK-8 assay, we found that the expression of circRNA_100782 is related to the proliferation of gastric cancer cells. Besides, we also found that circRNA_100782 regulated the migration ability of gastric cancer cells through transwell assay. The bioinformatics prediction and luciferase assay demonstrated that circRNA_100782 can serve as a molecular sponge to further regulate the expression of Rb by sponging with miR-574-3p; moreover, circRNA_100782 can serve as a ceRNA for miR-574-3p to further regulate the expression of Rb.
In this research, we discovered that circRNA_100782 was downregulated in gastric cancer cells and is associated with cell proliferation and invasion by inhibiting tumor suppressor gene Rb by interacting with miR-574-3p.
本研究旨在探讨胃癌组织中 circRNA_100782 的表达水平及其通过海绵吸附 miR-574-3p 的形式调节肿瘤抑制基因 Rb 的功能。
采用 qRT-PCR 检测不同阶段胃癌组织中 circRNA_100782 的表达。采用 CCK-8 法评估破骨细胞的增殖和分化。通过统计学分析检测 miR-574-3p 与 circRNA_100782 的相关性。通过生物信息学和荧光素酶报告基因实验检测 circRNA_100782 与 miR-574-3p 的相互作用和结合位点。将 Rb 的 3'-UTR 克隆到荧光素酶报告载体中，并构建 miR-574-3p 结合突变体以验证 miR-574-3p 对 Rb 表达的抑制调节作用。
与相邻的非癌性正常组织相比，人胃癌细胞中 circRNA_100782 和 Rb 的表达均下调。通过 qRT-PCR 和 CCK-8 法，我们发现 circRNA_100782 的表达与胃癌细胞的增殖有关。此外，我们还通过 Transwell 实验发现 circRNA_100782 通过调节胃癌细胞的迁移能力来调节胃癌细胞的迁移能力。生物信息学预测和荧光素酶报告基因实验表明，circRNA_100782 可以作为分子海绵，通过与 miR-574-3p 结合进一步调节 Rb 的表达；此外，circRNA_100782 可以作为 miR-574-3p 的 ceRNA 进一步调节 Rb 的表达。
在本研究中，我们发现 circRNA_100782 在胃癌细胞中下调，并通过与 miR-574-3p 相互作用抑制肿瘤抑制基因 Rb 从而与细胞增殖和侵袭有关。
