Research Paper Volume 8, Issue 11 pp 2948—2960

Figure 4. EWSAT1's oncogenic activity is in part through negative regulation of miR-326/330-5p, and then activation of cyclin D1 in NPC cells. (A) Up-regulated EWSAT1 in miR-326/330-5p treated CNE-1 and SUNE-1 cells, significantly reversed the growth-inhibitory role of miR-326/330-5p in NPC cells. (B) The 3'-UTR of cyclin D1 harbors two miR-326/330-5p cognate sites. (C) Relative luciferase activity of reporter plasmids carrying wild-type or mutant cyclin D1 3'-UTR in CNE-1 and SUNE-1 cells co-transfected with negative control (NC) or miR-326/330-5p mimic. (D) Statistical analysis of trypan blue staining. (E-F) Protein expression of cyclin D1 in miR-330-5p, miR-330-5p+EWSAT1, miR-326, miR-326+ewsat1, si-CCND1, EWSAT1, or EWSAT1+si-CCND1 treated CNE-1 and SUNE-1 cells. Assays were performed in triplicate. *P < 0.05. Means ± SEM are shown. Statistical analysis was conducted using student One-Way ANOVA test.