Abstract
Neuroblastoma, featured by a high rate of spontaneous remissions, is the most common extra-cranial solid tumor in infants and children. Numerous reports have demonstrated that MicroRNAs (miRNAs) play essential roles in cancer progression, including cell proliferation, apoptosis, invasion, metastasis and angiogenesis. miR-421 functions as an onco-miR in some malignancies. However, its role in neuroblastoma remains poorly understood. In the present study, we found that miR-421 was increased in neuroblastoma tissues compared with matched adjacent normal tissues. Forced overexpression of miR-421 substantially enhanced cell proliferation, cell-cycle progression, migration, and invasion of neuroblastoma cells. At the molecular level, tumor suppressor menin was found to be a target of miR-421. Furthermore, downregulation of menin by small interfering RNA oligos exhibited similar effects with overexpression of miR-421. On the other hand, overexpression of menin partially reversed the proliferative effects of miR-421 in neuroblastoma cells. Collectively, miR-421 may promote neuroblastoma cell growth and motility partially by targeting menin.
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Supplementary Figure 1
Expression levels of miR-421 were compared in SHSY5Y and SHEP cells by real-time PCR. (A)Expression of miR-421 was determined in SHEP cells after transfection of miR-421 mimics or negative controls (NC). (C) The cell viability of SHEP cells was determined by MTT assays. (D) The growth curve of SHEP cells after miR-421 transfection compared to NC. (E) The cell proliferative potential (BrdU) was determined in SHEP cells. A450 absorption was assayed after transfection for 24 hr. (F) The cell-cycle phase of SHSY5Y cells were analyzed by flow cytometry. *p < 0.05, ** p < 0.01, *** p < 0.001, compared with NC. (GIF 6 kb)
Supplementary Figure 2
(A) Expression levels of miR-421 were determined in three cell lines by real-time PCR. (B)Expression of miR-421 was determined in IMR-32 cells after transfection of miR-421 mimics or negative controls (NC). (C) The cell viability of IMR-32 cells was determined by MTT assays. (D) The growth curve of IMR-32 cells after miR-421 transfection compared to NC. (E) The cell proliferative potential (BrdU) was determined in IMR-32cells. A450 absorption was assayed after transfection for 24 hr. (F) The cell-cycle phase of IMR-32 cells was analyzed by flow cytometry. *p < 0.05, ** p < 0.01, *** p < 0.001, compared with NC. (GIF 6 kb)
Supplementary Figure 3
Representative Menin protein levels in human NB and adjacent normal tissues from two patients. 40 μg amount of tissue lysates were used in the western blot experiments. (GIF 0 kb)
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Li, Y., Li, W., Zhang, JG. et al. Downregulation of tumor suppressor menin by miR-421 promotes proliferation and migration of neuroblastoma. Tumor Biol. 35, 10011–10017 (2014). https://doi.org/10.1007/s13277-014-1921-1
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DOI: https://doi.org/10.1007/s13277-014-1921-1