microRNA-106b-mediated down-regulation ofC1orf24expression induces apoptosis and suppresses invasion of thyroid cancer

Abstract

We previously showed that C1orf24 expression is increased in thyroid carcinomas. Nonetheless, the mechanism underlying C1orf24 deregulation is not fully understood. It has been widely demonstrated that microRNAs are involved in post-transcriptional gene regulation in several diseases, including cancer. Using in silico prediction approach, five microRNAs that bind to the 3′-untranslated region (3′-UTR) of C1orf24 were identified. The expression of two selected microRNAs (miR-17-5p, miR-106b) and the expression of C1orf24 were tested in 48 benign and malignant thyroid lesions and in five thyroid carcinoma cell lines. miR-106b was down-regulated in thyroid cancer specimens and thyroid carcinoma cell lines, while C1orf24 expression was markedly increased. To demonstrate that miR-106b reduces C1orf24 expression, follicular (WRO) and papillary (TPC1) thyroid carcinoma cell lines were transiently transfected with miR-106b mimic. Ectopic expression of the miR-106b mimic significantly inhibits C1orf24 mRNA and protein expression in both WRO and TPC1 cells. Dual-luciferase report assays demonstrated that miR-106b directly targets C1orf24 by binding its 3′- UTR. Moreover, miR-106b-mediated down-regulation of C1orf24 expression increased apoptosis and inhibited migration. We additionally demonstrated that siRNA against C1orf24 significantly decreased its expression, inhibited cell migration and cell cycle progression while induced apoptosis. In summary, our findings not only provide new insights into molecular mechanism associated with C1orf24 overexpression in thyroid carcinomas but also show that C1orf24 might increase proliferation and cell migration. Thus, decreasing C1orf24 levels, by restoring miR-106b function, may have therapeutic implications.

MiR‐106b regulates the apoptosis and tumorigenesis of hepatocellular carcinoma via targeting Zinc finger and BTB domain‐containing protein 7A (Zbtb7a)

“…It also has been reported that miR‐106b targets tumor suppressor gene SET domain containing protein 2 (SETD2) to regulate the proliferation of tumor cells in renal cell carcinoma . Moreover, miR‐106b was also reported to mediate downregulation of C1orf24 expression, which increased apoptosis and inhibited migration in thyroid carcinoma . However, it is not clear which role, tumor suppressor or promoter, does miR‐106b act as in HCC.…”

RASSF10 is Epigenetically Inactivated and Suppresses Cell Proliferation and Induces Cell Apoptosis by Activating the p53 Signalling Pathway in Papillary Thyroid Carcinoma Cancer

“…Next, RNA was reversed transcribed into cDNA using SuperScript III (Invitrogen) and used as the template for real time-PCR (RT-PCR) assays. The conditions of RT-PCR were performed as previously described [24]. The sequences of the primers used in this experiment are listed in Table 1.…”

PVALB diminishes [Ca2+] and alters mitochondrial features in follicular thyroid carcinoma cells through AKT/GSK3β pathway

“…Antibody against PVALB (1:1000, Sigma-Aldrich) was used to select the cell line with lower basal expression of PVALB. Western blot analysis was performed as described previously (Latini et al 2008, Carvalheira et al 2015. WRO cells were simultaneously incubated for 40 min with 5 μM Fluo-4 AM (cytoplasmic) and Rhod-2 AM (mitochondrial) Ca 2+ fluorescent indicators at 37°C (Molecular Probes).…”

“…Protein extraction and quantification were performed in transfected WRO and FTC-133 cells, as described previously (Latini et al 2008, Carvalheira et al 2015. Briefly, cells were lysed in 50 mM Tris-HCl, 100 mM NaCl, 50 mM NaF, 1 mM NaVO 4 and 0.5% Triton-X solution with protease inhibitors (Complete Cocktail, Roche, Basel, Switzerland).…”

“…The signal was detected by ImageQuant LAS 4000 system (GE Healthcare). The bands were quantified using Image-J Software, as described previously (Carvalheira et al 2015).…”