Figure 1

miR-133b targets NUP214 and their expressions are inversely related. (a) Expression of endogenous miR-133b in different cancer cell lines compared to control oral epithelial cells. (b) Corresponding levels of endogenous NUP214 in the same cell lines compared to control oral epithelial cells. (c) Reciprocal relation of miR-133b and NUP214 expressions in HNSCC tissues. Relative expression values were calculated and the data plotted as shown. ‘n’ represents number of tumour samples; asterisk denotes significant difference of expression in tumour versus normal samples with P-value < 0.0005. (d) NUP214 is reduced at the mRNA level in presence of ectopic miR-133b. UPCI:SCC084 cells were transiently transfected with 0, 0.25, 0.5, 1 and 2 μg of pSB-miR-133b. (e) Representative picture showing Nup214 protein level decreases upon ectopic miR-133b expression. UPCI:SCC084 cells were transiently transfected with 0, 0.25, 0.5, 1 and 2 μg of pSB-miR-133b. Cell lysates were prepared followed by Western blot with antibodies against Nup214 and β-actin. For (a) and (c), total RNA from oral swab (control), UPCI:SCC084, SCC25, SW480, HCT116 cells and HNSCC tumour and normal tissues were reverse transcribed using miR-133b-specific stem-loop primers. cDNAs were subjected to RT-PCR. Relative expression values were normalized to those of miR-17-5p or (for cell lines) or U6 snRNA (for tissues). For (b), (c), and (d), cDNA was subjected to RT-PCR using NUP214-specific primers. Relative expression values were normalized to those of GAPDH (for cell lines) or 18 s rRNA (for tissues). For (a), (b) and (d), data represent three independent experiments and are shown as average ± standard deviation (S.D). For (e) images are representative of three independent experiments. For all figures, asterisk denotes significant change when compared to either control or miR-133b-treated condition with p value <0.05; NS – not significant.