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: Our previous study established radioresistance mRNA profiles and KEGG pathway analysis showed upregulated mRNA might function via the human T-cell leukemia virus type I (HTLV-I) infection pathway (which contains the most upregulated genes).(17) MiRNA target prediction program RNAhybrid indicated three upregulated genes (i.e. WNT2B, ICAM1 and HLA-F) in the HTLV-I infection pathway were targeted by miR-185-3p (Fig. 4a). Herein, mRNA expression of WNT2B, ICAM1 and HLA-F were detected in NPC cells transfected with the miR-185-3p mimic/inhibitor. Interestingly, only WNT2B mRNA demonstrated converse alterations (Fig. 4b). Thus, further validation on the relationship between WNT2B and miR-185-3p was performed. Western blotting analyses showed that in miR-185-3p-mimic transfected cells, WNT2B decreased and downstream β-catenin and p-GSK-3β were also altered (Fig. 4c). In addition, we detected the expression of miR-185-3p and WNT2B in 15 NPC tissues and six NCET samples via qRT-PCR. Our results confirmed that the average expression level of miR-185-3p was significantly lower, but WNT2B was obviously upregulated in NPC specimens compared with NCET tissue samples (Fig. 4d). A negative correlation between miR-185-3p and WNT2B was also observed (R = −0.631; P = 0.002; Fig. 4e). These results revealed that WNT2B might be the target gene of miR-185-3p.
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.