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Updated: Dec 13, 2025

Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells
Published on: May 9, 2020
Identification and Expression of Several Circular RNAs and Knockdown of hsa_circ_0005556 Exerts Oncogenic Functions
Hanke Geng1, Kaixuan Li1, Qi Pan2
1Clinical Laboratory, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi, China (mainland).
Abstract:
BACKGROUND Gastric cancer (GC) remains one of the most fatal digestive cancers in the world; nevertheless, its etiology remains vague. With the development of bioinformatics analysis, numerous circular RNAs (circRNAs) have been found to be dysregulated in GC. However, the functions of a large portion of dysregulated circRNAs in GC need further validation. In this study, we aimed to validate the biological functions of circ_0005556, which was previously identified to be dysregulated in GC. MATERIAL AND METHODS Levels of circRNAs and miRNAs in GC tissues and cells were estimated by qRT-PCR. The target miRNAs of circ_0005556 were predicted by bioinformatics methods. The interplay between circ_0005556 and miR-767-5p was validated by dual-luciferase reporter and circRNA immunoprecipitation assays. The effects of circ_0005556 and miR-767-5p on GC cell viability, apoptosis, migration, and invasion were assessed by MTT, flow cytometry, wound-healing and in vitro transwell experiments, respectively. RESULTS The upregulation of circ_0005556 was validated by qRT-PCR in GC tissues and cells, and a higher circ_0005556 level indicated a poorer prognosis. miR-767-5p was demonstrated to target circ_0005556 in GC cells, and a negative correlation was found between their expression levels in GC tissues. Knockdown of circ_0005556 promoted miR-767-5p expression in GC cells. Knockdown of circ_0005556 was revealed to repress GC cell viability, invasion, and migration and to promote GC cell apoptosis. Moreover, overexpression of miR-767-5p could significantly augment the repressive impacts of circ_0005556 knockdown on GC cell progression in vitro. CONCLUSIONS The in vitro knockdown of circ_0005556 remarkably repressed GC cell progression by increasing the expression of miR-767-5p.
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