RGS16 expression is decreased in chondrosarcoma and RGS16 transfection counteracts the effects of miR-181a on VEGF and MMP1 expression. RGS16 expression.

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RGS16 expression is decreased in chondrosarcoma and RGS16 transfection counteracts the effects of miR-181a on VEGF and MMP1 expression. RGS16 expression is decreased in chondrosarcoma and RGS16 transfection counteracts the effects of miR-181a on VEGF and MMP1 expression. RGS16 was evaluated with real-time PCR and Western blotting; VEGF and MMP1 were quantitated with ELISA, as described in Materials and Methods. A, articular cartilage and HCS; B, chondrosarcoma cell lines compared with chondrocytes; C, xenograft tumors compared with cell lines in vitro. NCL, articular cartilage (N = 12); CSI, HCS grade 1 (N = 7); CSII-III, HCS grades 2 and 3 (N = 16); CH, chondrocytes; JJ, CS-1, chondrosarcoma cell lines (CS-1 derived from grade 3 tumor); n, normoxia; h, hypoxia (2% O2); JJ-tumor, CS-1-tumor, xenograft tumors. Levels of mRNA are shown as mean ± SD, N = 3. CS-1 cells were transfected with control, RGS16 cDNA construct, miR-181a mimic, or both RGS16 cDNA construct and miR-181a, as described in Materials and Methods. VEGF (D) and MMP1 (E) in CM after 2 days in hypoxic culture; #, P < 0.05; *, <0.001; **, P < 0.01. F, Western blot analysis confirming the effect of RGS16 transfection and miR-181a on RGS16 protein levels. G, schematic representation illustrating the role of miR-181a in chondrosarcoma angiogenesis and metastasis. Xiaojuan Sun et al. Mol Cancer Res 2015;13:1347-1357 ©2015 by American Association for Cancer Research