Aberrant expression of miR-10b has been described in many cancers but remains unexplored in nasopharyngeal carcinoma (NPC). Therefore, we aimed to study the miR-10b expression level in 43 NPC biopsies collected from Tunisian patients and three NPC xenografts. Then, we investigated the correlation between miR-10b expression and its upstream regulators LMP1/Twist1 as well as its adjacent gene HoxD4. We showed that miR-10b was significantly up-regulated in NPC biopsies compared to non-tumor nasopharyngeal tissues (fold change 153; p = 0.004) and associated with advanced clinical stage and young age at diagnosis (p = 0.005 and p = 0.011, respectively). In addition, over-expression of miR-10b was positively associated with the transcription factor Twist1 as well as the EBV oncoprotein LMP1 (fold change 6.32; p = 0.014, fold change 6.58; p = 0.01 respectively). Furthermore, higher level of miR-10b was observed in tumors with simultaneous expression of LMP1 and Twist1, compared to those expressing only Twist1 (fold change 2.49; p = 0.033). Meanwhile, the analysis of the link between miR-10b and its neighbor gene HoxD4 did not show any significant correlation (Fisher test p = 0.205; Mann–Whitney test p = 0.676). This study reports the first evidence of miR-10b over-expression in NPC patients. Furthermore, our findings can support hsa-miR-10b gene regulation through LMP1/Twist1 in NPC malignancy.
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This work was supported by a grant of the Tunisian Ministry of High Education and Scientific Research. We wish to thank Pr Adnane Hammami for allowing us to perform the RT-q-PCR experiments in the microbiology laboratory at the School of Medicine of University of Sfax.
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Generating a standard curve to assess reaction efficiency. A Amplification curves and their corresponding melt curves of qPCR reactions using a series of tenfold dilutions of specific cDNA (1, 1/10, 1/100, 1/1000, and 1/10000) of miR-10b (A1) and U6 (A2). B Standard curve with the Ct plotted against the log of the starting quantity of template for each dilution. The equation of the generated curve is shown above the graph. The calculated efficiency was 103 and 101 % for miR10b and U6, respectively (TIFF 2886 kb)
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Allaya, N., Khabir, A., Sallemi-Boudawara, T. et al. Over-expression of miR-10b in NPC patients: correlation with LMP1 and Twist1. Tumor Biol. 36, 3807–3814 (2015). https://doi.org/10.1007/s13277-014-3022-6
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