The Effect of Metformin on the Proliferation, Apoptosis and CD133 mRNA Expression of Colon Cancer Stem Cells by Upregulation of miR 342-3p.

OBJECTIVE

To explore whether metformin (MET) can affect the biological behaviour and CD133 mRNA expression of CD133 colon cancer stem cells (CCSCs) through miR-342-3p.

METHODS

The direct immunomagnetic bead method was used to select CD133 CCSCs from the SW480 and HCT116 cell lines, and miRNA-tailing qRT-PCR was used to detect the expression changes of tumor suppressor-related miRNAs (miR-34a, miR-126, miR-143, miR-145, miR-342-3p, miR-342-5p) after MET intervention. Then, miR-342-3p with markedly significant differential expression was selected as the target miRNA. The lentiviruses LV16-hsa-miR-342-3p inhibitor and LV16-NC were used for the transfection inhibition test. CCK-8, flow cytometry, and qRT-PCR were used to detect the cell viability, apoptosis rate, and CD133 mRNA expression of CD133 CCSCs.

RESULTS

Under the high-glucose environment, the expression of tumor suppressor-related miRNAs in CCSCs changed differently ( <0.05), MET also had different effects on the expression of tumor suppressor-related miRNA under different glucose concentrations (<0.05). Among them, MET upregulates the expression of miR-342-3p in CCSCs for the first time. The results of the lentiviruses transfection inhibition test showed that after miR-342-3p was inhibited, the cell viability and apoptosis rate of CD133 CCSCs did not change significantly compared with before inhibition (>0.05), but the expression of CD133 mRNA markedly increased (<0.05). Meanwhile, after MET intervention, the apoptosis rate and the expression of CD133 mRNA of CD133 CCSCs was significantly increased, and the proliferation of CD133 CCSCs was obviously inhibited (<0.05).

CONCLUSION

MET upregulating the expression of miR-342-3p may not have a significant effect on the proliferation and apoptosis of CD133 CCSCs, but it can reduce the expression of CD133 mRNA in CD133 CCSCs.