Beta-actin was used as a loading control. Images are representative of three independent selleck compound experiments. B shows MMPs protein levels (expressed as percentages of controls) (n = 3). Numbers in the box represent the concentration of risedronate in μM added to the cells. Bars represent MMPs protein levels (expressed as percentages of controls)
of each band ± standard deviation. Risedronate suppressed MMP-2 and MMP-9 mRNA levels in both cell lines RT-PCR was used to determine whether risedronate suppresses MMP-2 and MMP-9 at the transcription levels. Risedronate was found to attenuate MMP-2 and MMP-9 mRNA levels dose-dependent in both cell lines (p < 0.05) (Fig. 6). Figure 6 Risedronate suppressed the expressions of MMP-2 and MMP-9 mRNA in SaOS-2 and U2OS cells. this website (A) Cells were treated with the indicated concentrations of risedronate for 48 h and then processed for RT-PCR. Beta-actin was used as a loading control. Images are representative of three independent experiments. MMPs mRNA levers (expressed as percentages of controls) are shown in B (n = 3). Numbers in the box represent the concentration of risedronate in μM added to the cells. Bars represent MMPs mRNA levels (expressed as percentages of controls) of each band ± standard deviation. Discussion Osteosarcoma is an aggressive malignant bone disorder exerting a high
potential to invade and Fenbendazole metastasize. A number of studies have demonstrated the beneficial effects of bisphosphonates on bone metastases from different solid tumors, such as, those of the breast, prostate and renal cell carcinoma [29, 30]. In the majority of previous studies, first or second-generation bisphosphonates have been examined at the relatively high concentrations required to inhibit the cell proliferation of osteosarcoma
cells [31, 32]. In addition, third-generation bisphosphonates have been reported to induce osteosarcoma cell apoptosis. Evdokiou and colleagues studied the third-generation bisphosphonate, zoledronic acid (ZOL), and found that it dose- and time-dependently decreased cell proliferation in a panel of human osteosarcoma cell lines , Tadahiko Kubo and Shoji Shimose reported that minodronate and incadronate perturb the cell cycle and induce the apoptosis of SaOS-2 cells . However, the molecular mechanism underlying inhibition by BPs has not been determined. Cheng YY et al. reported that alendronate reduces MMP-2 secretion and induces tumor cell apoptosis in osteosarcoma , but the molecular targets and modes of action of MMP-2 and MMP-9 inhibitors, like risedronate, are substantially unknown. In the present study, we found that risedronate suppresses cell invasion and the gelatinolytic SB203580 manufacturer activities and protein and mRNA expressions of MMP-2 and MMP-9 in the SaOS-2 and U2OS osteosarcoma cell lines.