Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: MiR-185-3p and miR-324-3p Predict Radiosensitivity of Nasopharyngeal Carcinoma and Modulate Cancer Cell Growth and Apoptosis by Targeting SMAD7

doi: 10.12659/MSM.895660

Figure Lengend Snippet: miR-185-3p and miR-324-3p can target 3′UTR of SMAD7 and modulate growth and apoptosis of NPC cells. ( A ) Predicted binding sites between miR-185-3p/miR-324-3p and 3′UTR of SMAD7. ( B ) CNE-2 cells were transfected with 50 nM miR-185-3p or miR-324-3p mimics for overexpression. ( C ) qRT-PCR analysis of SMAD7 expression at both mRNA (upper panel) and protein level (lower panel) in CNE-2 cells with miR-185-3p or miR-324-3p overexpression. ( D, E ) Dual luciferase assay of relative firefly luciferase activity in CNE-2 cells co-transfected with miR-183-3p ( D ) or miR-324-3p ( E ) and the reconstructed plasmids carrying wild-type or mutant miR-185-3p or miR-324-3p binding sequence. ( F, G ) CCK-8 assay of relative cell proliferation in CNE-2 cells with only miR-185-3p or miR-324-3p overexpression ( F ) or the cells with SMAD7 overexpression or combined SMAD7 and miR-185-3p or miR-324-3p overexpression ( G ). ( H ) Representative images of flow cytometry analysis of apoptotic CNE-2 cells with only miR-185-3p or miR-324-3p overexpression or the cells with SMAD7 overexpression or combined SMAD7 and miR-185-3p or miR-324-3p overexpression. *, ** and *** donate significance at 0.05, 0.01 and 0.001 levels, respectively.

Article Snippet: Human undifferentiated non-keratinizing NPC cell line CNE-2 was obtained from the Experiment Animal Center of Sun Yat-Sen University and HEK-293T cells were obtained from ATCC.

Techniques: Binding Assay, Transfection, Over Expression, Quantitative RT-PCR, Expressing, Luciferase, Activity Assay, Mutagenesis, Sequencing, CCK-8 Assay, Flow Cytometry