OBJECTIVE: The aim of this study was to explore the effect of micro ribonucleic acid (miR)-200c on the proliferation and apoptosis of Wilms tumor cells, and to further elucidate its potential mechanisms.
PATIENTS AND METHODS: Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was used to detect the expression level of miR-200c in cancer tissues and adjacent normal tissues of 20 patients with Wilms tumor. Human primary Wilms tumor cells were taken as research objects, and were divided into Control group and miR-200c mimic group. In miR-200c mimic group, miR-200c was overexpressed in Wilms tumor cells using liposome transfection technology. Subsequently, the proliferation and apoptosis of cells in each group were observed by functional assays. The expression levels of phosphorylated protein kinase B (p-Akt), total Akt (T-Akt) and glucose transporter protein type 1 (GLUT1) in each group of cells were finally detected by Western blotting.
RESULTS: In Wilms tumor patients, the expression level of miR-200c in cancer tissues was notably lower than that in adjacent normal tissues (p<0.05). Wilms tumor cells were cultured in vitro, and were transfected with miR-200c mimic. Subsequent cell counting kit-8 (CCK-8) assay results showed that the proliferation ability of cells in miR-200c mimic group was remarkably weakened (p<0.05). Colony formation assay indicated the number of formed colonies in miR-200c mimic group was remarkably less than that in Control group (p<0.05). Western blotting results manifested that overexpression of miR-200c markedly increased the ratio of B-cell lymphoma protein 2 (Bcl-2) to Bcl-2-associated X protein (Bax) (p<0.05). Flow cytometry results revealed that miR-200c overexpression significantly elevated the apoptosis rate of Wilms tumor cells (p<0.05). In addition, it was discovered that the overexpression of miR-200c could prominently reduce the phosphorylation level of intracellular Akt and the expression of its downstream protein GLUT1. Finally, immunohistochemical staining results verified that the expression levels of p-Akt and GLUT1 in cancer tissues of Wilms tumor patients were significantly higher than those in adjacent normal tissues (p<0.05).
CONCLUSIONS: MiR-200c was lowly expressed in cancer tissues of Wilms tumor patients. Besides, overexpression of miR-200c inhibited the proliferation and promoted the apoptosis of cells through targeted inhibition of the Akt/GLUT1 signaling pathway.Free PDF Download
To cite this article
G.-Z. Zhao, Y.-Q. Niu, Z.-M. Li, D. Kou, L. Zhang
MiR-200c inhibits proliferation and promotes apoptosis of Wilms tumor cells by regulating akt signaling pathway
Eur Rev Med Pharmacol Sci
Vol. 24 - N. 12