Knockdown of circ_0003340 induces cell apoptosis, inhibits invasion and proliferation through miR-564/TPX2 in esophageal cancer cells.

Circular RNA (circRNA) is a promising biomarker of cancer occurrence and development. The different expression levels of circRNAs in various cancers also make them possible therapeutic targets. In this work, we researched the function and underlying mechanisms of circ_0003340 (circ3340) in esophageal cancer EC1 and EC9706 cells. Firstly, we found the expression levels of circ3340 are higher in ESCC and two esophageal cancer cells than in adjacent normal tissues and Het-1a cells. Bioinformatics analysis showed circ3340 has a binding site with miR-564. This was verified by luciferase assay, which revealed that miR-564 can be sponged by circ3340, and that the TPX2 3'UTR is a direct target of miR-564. Upregulation of miR-564 decreased TPX2 protein levels, as shown by Western blot. Moreover, knockdown of circ3340 or enhancement of miR-564 expression had similar effects in EC1 and EC9706 cells, i.e., inducing cell apoptosis, inhibiting cell proliferation, and arresting cell invasion. Downregulation of circ3340 had a negative influence on EC1 and EC9706 cells by affecting the miR-564/TPX2 pathway. Additionally, animal experiments revealed that downregulation of circ3340 inhibited tumor growth in vivo, making circ3340 a potential therapeutic target for patients with esophageal squamous cell cancer.

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