Effects of MDM4 Gene Regulation by miR-34a on LOVO Cell Proliferation.

This study aims to investigate the effects of regulation of murine double minute protein4 (MDM4) expression by microRNA (miR)-34a on the proliferation of colorectal cancer LOVO cells. Prediction results obtained using a gene microarray showed that MDM4 is a specific miR-34a target gene. The interaction between miR-34a and the 3'-untranslated region (UTR) of MDM4 was detected using a luciferase reporter system. The effect of miR-34a on MDM4 protein expression was detected by Western blotting, and the effect of miR-34a transfection on LOVO cell proliferation was detected using an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and flow cytometry. The luciferase activity in LOVO cells after addition of pmirGLO-UTR+miR-34a mimic was only 44% of that obtained with pmirGLO-UTR+miR-34a (p<0.01), and the luciferase activity in LOVO cells after addition of pmir-GLO-mutUTR+miR-34a was 94% of that observed after pmirGLO-UTR+miR-34a addition (p=0.57). These results indicated that miR-34a interacted with the 3'-UTR of MDM4. Results of Western blotting revealed that MDM4 protein expression level in LOVO cells after addition of miR-34a mimic was 29% that of non-treated LOVO cells (p<0.05), indicating that miR-34a negatively regulates MDM4 protein expression. The growth rate of LOVO cells with miR-34a overexpression was significantly reduced compared with that of non-treated LOVO cells (p<0.05), indicating that miR-34a overexpression inhibits LOVO cell proliferation. miR-34a negatively regulates MDM4 gene expression to inhibit LOVO cell proliferation. © 2020 by the Association of Clinical Scientists, Inc.

Citation

Shanshan Li, Yang Pei, Wei Zhang, Liying Sun, Miao Jiang, Guoyin Shang, Sijia Niu, Zhibin Ma. Effects of MDM4 Gene Regulation by miR-34a on LOVO Cell Proliferation. Annals of clinical and laboratory science. 2020 May;50(3):342-347

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PMID: 32581023

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