Anti-BIRC7 monoclonal antibody, clone 62.4_3B4

Osteoblasts and osteoclasts coordinate to monitor the dynamic osteogenic balance between bone formation and bone resorption. Hence, an investigation of the regulatory mechanisms underlying osteogenic osteoblast differentiation will provide more methods for bone repair and bone regeneration. In the present study, human osteoblast hFOB 1.19 cells were cultured. MicroRNA-214 (miR-214) expression significantly down-regulated during the osteogenic differentiation of hFOB 1.19 cells. In addition, miR-214 overexpression by miR-214 precursor transfection markedly inhibited the expression of alkaline phosphatase (ALP), collagen type I alpha 1 (col1 alpha 1) and runt-related transcription factor 2 (Runx2), which concomitantly decreased ALP activity and the number of mineralized nodules but promoted the expression of signal transducer and activator of transcription 1 (STAT1), an osteogenesis blocker. We next found that miR-214 inhibited the expression of baculoviral IAP repeat-containing 7 (BIRC7), a member of the inhibitor of apoptosis proteins family. However, BIRC7 overexpression, which was induced by plasmid transfection, notably reversed the inhibitory effects of miR-214, indicating a potential BIRC7-dependent osteogenic differentiation manner mediated by miR-214. Taken together, our results demonstrate for the first time that miR-214 suppresses osteogenesis by targeting BIRC7, providing a possible therapeutic target for bone degenerative diseases.

Neuroblastoma (NB) is the most common extra-cranial solid tumor in childhood with the overall 5 years' survival less than 40%. Polo-like kinase 1 (PLK1) is a serine/threonine-protein kinase expressed during mitosis and over expressed in multiple cancers, including neuroblastoma. We found that higher PLK1 expression related to poor outcome of NB patients. BI2536, a small molecule inhibitor against PLK1, significantly reduced cell viability in a panel of NB cell lines, with IC50 less than 100 nM. PLK1 inhibition by BI 2536 treatment induced cell cycle arrest at G(2)/M phase and cell apoptosis in NB cells. Realtime PCR array revealed the PLK1 inhibition related genes, such as BIRC7, TNFSF10, LGALS1 and DAD1 et al. Moreover, autophagy activity was investigated in the NB cells treated with BI 2536. BI 2536 treatment in NB cells increased LC3-1I puncta formation and LC3-II expression. Formation of autophagosome induced by BI 2536 was observed by transmission electron microscopy. However, BI2536 abrogated the autophagic flux in NB cells by reducing SQSTM1/p62 expression and AMPK alpha(T172) phosphorylation. These results provide new clues for the molecular mechanism of cell death induced by BI 2536 and suggest that BI 2536 may act as new candidate drug for neuroblastoma.