Up-Regulation of miR-96-5p Inhibits the Proliferation of FaDu Cell Line by Targeting mTOR

Abstract

Hypopharyngeal carcinoma is an upper respiratory-gastrointestinal tract cancer, with a complicated pathogenesis and high mortality. We predicted that hsa-miR-96-5p targeted mTOR. To test this hypothesis, the overexpression vector of miR-96-5p, mTOR 3'UTR and its mutant vector with cobinding sites of miR-96-5p, and shRNA-mTOR vectors were constructed. Two groups of FaDu cells were transfected with and without metformin (10 mmol/L), and cultivated for 48, 72 and 96 hours. RTPCR and Western blotting experiments showed that upregulation of miR-96-5p inhibited proliferation of FaDu cells. At the same time, we found that upregulation of miR-96-5p inhibited the growth of FaDu cells treated with metformin. After detecting the mTOR mutant 3'UTR loci by dual luciferase assay, we found that the recorded fluorescence values were much higher than that of the wildtype. We also determined that the ratio of firefly fluorescence value/Renilla fluorescence value was significantly higher than that of wild-type, indicating that mTOR was the direct target of the miR-965-p gene. In conclusion, miR-96-5p inhibited proliferation of FaDu cells by targeting mTOR, and it may serve as a potential target for tumor growth inhibition.