miR-185-3p regulates nasopharyngeal carcinoma radioresistance by targeting WNT2B in vitro.
Bottom Line: Luciferase reporter assays confirmed that miR-185-3p directly targeted the coding region of WNT2B.Furthermore, we found radioresistance decreased in WNT2B-silenced NPC cells.We concluded that miR-185-3p contributed to the radioresistance of NPC via modulation of WNT2B expression in vitro.
Affiliation: Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China.Show MeSH
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Mentions: As we predicted, WNT2B is most likely the target gene of miR-185-3p. A series of experiments was performed to confirm our prediction. Hybridization of miR-185-3p and WNT2B mRNA can be predicted using RNAhybrid software and the minimum free energy required for this hybridization is −35.2 kcal/mol (Fig. 5a). Thus, specific targeting of WNT2B by miR-185-3p was examined using luciferase reporter assays. A mutant WNT2B reporter gene was constructed by deleting the seed sequence GGT GCG GAG GAA GCU GCG CAG CUC CC and mutating this sequence to GCT GGG CTC CTT TCT CGG GTC GGG GC (Fig. 5b). Our data revealed that disruption of the binding sites between miR-185-3p and the coding region of WNT2B mRNA abolished the miR-185-3p-mediated inhibition of WNT2B luciferase activity (P < 0.01; Fig. 5b). Taken together, these data indicate that miR-185-3p inhibits the expression of WNT2B protein via specific binding to the coding region of its mRNA.
Affiliation: Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China.