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Interleukin-6 antisense deoxyoligonucleotides inhibit bone resorption by giant cells from human giant cell tumors of
S V Reddy1, S Takahashi, M Dallas
1Department of Medicine/Hematology, University of Texas Health Science Center, San Antonio.
Abstract:
The effects of antisense constructs to IL-6 on the bone-resorbing capacity of purified giant cells from giant cell tumors of bone were examined to further define the role of IL-6 in human osteoclastic bone resorption. In addition, we wanted to determine the utility of antisense constructs to cytokines produced by osteoclasts as probes to identify the molecular events responsible for the bone-resorptive process. Giant cells were cultured on sperm whale dentin for 24 h in the presence of fluoresceinated antisense or scrambled antisense deoxyoligonucleotides complementary to IL-6 mRNA. The giant cells actively incorporated these oligonucleotides, as evidenced by their intense fluorescence. The number of resorptive lacunae formed and the area of the dentin resorbed were significantly decreased in cultures of giant cells treated with antisense IL-6 constructs compared with control cultures treated with scrambled antisense constructs to IL-6 (60 +/- 13 versus 12 +/- 6 lacunae and 1.2 +/- 0.3 versus 0.26 +/- 0.1 x 10(5) microns2). IL-6 levels in conditioned media from giant cell cultures treated with IL-6 antisense constructs were fourfold lower than those in control media obtained from giant cells treated with scrambled antisense constructs to IL-6. These data confirm the capacity of IL-6 antisense oligomers to block IL-6 production by these cells. These observations show that IL-6 plays an important role in the bone-resorptive process of human osteoclasts and suggest that antisense constructs to cytokines produced by bone cells may be useful for determining the molecular events occurring during bone resorption.
Insights
Antisense constructs targeting interleukin-6 (IL-6) significantly reduced bone resorption by giant cells from bone tumors. This confirms IL-6
Area of Science:
- Bone biology
- Molecular biology
- Oncology
Background:
- Giant cell tumors of bone are characterized by numerous multinucleated giant cells.
- Osteoclasts, including those in giant cell tumors, are responsible for bone resorption.
- Interleukin-6 (IL-6) is a cytokine implicated in various cellular processes, including bone metabolism.
Purpose of the Study:
- To investigate the role of IL-6 in the bone-resorbing activity of human osteoclasts.
- To assess the effectiveness of antisense technology in targeting cytokine production by osteoclasts.
- To identify molecular mechanisms underlying osteoclastic bone resorption.
Main Methods:
- Purified giant cells from bone tumors were cultured on dentin.
- Cells were treated with fluoresceinated antisense or scrambled antisense deoxyoligonucleotides targeting IL-6 mRNA.
- Bone resorption was quantified by measuring resorptive lacunae and resorbed dentin area.
- IL-6 levels in conditioned media were measured.
Main Results:
- Giant cells actively incorporated the deoxyoligonucleotides.
- Treatment with IL-6 antisense constructs significantly decreased the number and area of resorptive lacunae compared to controls.
- IL-6 levels in conditioned media were significantly lower in cells treated with IL-6 antisense constructs.
- Antisense oligomers effectively blocked IL-6 production.
Conclusions:
- IL-6 plays a critical role in the bone-resorptive capacity of human osteoclasts.
- Antisense constructs targeting cytokines are effective tools for studying bone resorption.
- This research provides insights into the molecular mechanisms of bone resorption in giant cell tumors of bone.