Fig. 1
TSPAN15 is overexpressed in OSCC and negatively regulated by miR-339. a Relative expression levels of TSPAN15 detected by qRT-PCR in 46 OSCC tissues compared with their adjacent non-tumor tissues. Expression of β-actin was used as an endogenous control. **P < 0.001, paired Student’s t-test. b qRT-PCR and western blot analysis of TSPAN15 expression in one immortalized oesophagus cell line (NE1) and ten OSCC cell lines. Expression of β-actin was used as an endogenous control. The data are represented as the mean ± s.d. of three independent experiments. *P < 0.01, **P < 0.001, Student’s t-test. c Representative of TSPAN15 expression in normal epithelium and OSCC tumor tissues detected by immunostaining with anti-TSPAN15 antibody (brown). The slide was counterstained with hematoxylin. Scale bar, 50 µm. d Kaplan−Meier analysis revealed that TSPAN15 overexpression was related with poorer overall survival of OSCC patients. P < 0.001, log-rank test. e Higher expression levels of miR-339 detected by qRT-PCR in 46 OSCC tissues compared with their adjacent non-tumor tissues. **P < 0.001, paired Student’s t-test. f Analysis showing linear regressions and significant Pearson correlations of miR-339-5p with TSPAN15 in 46 pairs of OSCC samples. g The mRNA levels of TSPAN15 decreased when miR-339-5p mimics was transfected into KYSE30 and KYSE140 cells compared with control cells. The data are represented as the mean ± s.d. of three independent experiments. **P < 0.001, Student’s t-test. h Luciferase assay confirmed that miR-339-5p targeted the 3′-UTR region of TSPAN15. MiR-339-5p-expressing cells and control cells were co-transfected with wt-T15-luc, mu-T15-luc or empty vector with pRL-TK, and relative luciferase activity was detected. The data are represented as the mean ± s.d. of three independent experiments. *P < 0.01, Student’s t-test
