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Updated: Jan 5, 2026

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Downregulated miR-585-3p promotes cell growth and proliferation in colon cancer by upregulating PSME3
Chunmei Liu1,2, Juan Yang1,2, Han Wu1,2
1Department of Pathology, Luohe Central Hospital, Luohe 462000, People's Republic of China.
Background:
Upregulation of PSME3 and its oncogenic roles have been reported in colon cancer recently. However, the underlying mechanism of PSME3 upregulation remains unknown. Here, we explored the expression of PSME3 and subsequently uncovered its mechanism in colon cancer.
Materials And Methods:
The expression of PSME3 was analyzed by using online databases, Oncomine and UALCAN. qPCR was carried out to detect the expression of PSME3 in collected colon cancer tissues and cell lines. Moreover, the promoter methylation and the hnRNA level of PSME3 were also analyzed by online database and qPCR, respectively. The candidate miRNAs targeting PSME3 were predicted by Starbase 3.0 and validated by luciferase reporter system. CCK-8, plate colon formation, and Edu incorporation were applied to study the functions of miRNA in colon cancer. The expression of miRNA and its correlation with PSME3 were detected in colon cancer tissues.
Results:
Oncomine and UALCAN data indicate PSME3 is obviously upregulated in colon cancer tissue samples which is further confirmed in collected colon cancer tissues and cells by qPCR. No significant difference in methylation status promoter of PSME3 was observed between colon and colon cancer tissues. The hnRNA level of PSME3 was comparable between colon epithelial cell and colon cancer cells. miR-585-3p is predicted to directly target PSME3 and is validated by luciferase reporter assay. Then, miR-585-3p downregulation is confirmed and miR-585-3p restoration can suppress cell growth and proliferation by inhibiting PSME3 in colon cancer indicating by CCK-8, plate colon formation, and Edu incorporation. Moreover, negative correlation in expression between miR-585-3p and PSME3 was observed in our collected tissues samples.
Conclusion:
We reveal for the first time that miR-585-3p downregulation accounts for the overexpression of PSME3 in colon cancer. Moreover, miR-585-3p, serving as a tumor suppressor, can inhibit cell growth and proliferation in colon cancer by targeting PSME3.
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