| Effects of RANKL on Breast Cancer Cell Lines |
| 유방암 세포주에 대한 RANKL의 효과에 관한 연구 |
| 정호연,황창선1,강영순1,문인걸1,임창훈,한기옥,장학철,박원근,윤현구,한인권 |
| 성균관대학교 의과대학 삼성제일병원 내과, 내분비연구실1 |
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| Abstract |
Background
Breast cancer metastasis commonly induces osteolytic bone lesions. This osteolytic metastasis is mediated by the differentiation and activation of osteoclasts through upregulation of RANKL. Breast cancer cells with the capacity to stimulate osteoclastic bone resorption may enrich the bone microenvironment with growth factors that may alter behavior of tumor cells. Therefore, we investigated whether the locally increased RANKL can affect the invasion of breast cancer or have the influence on the change of gene expressions in breast cancer cells.
Methods
We assessed NF-B activation using electrophoretic mobility shift assay in RANKL treated breast cancer cell lines (MCF-7 & MDA-MB-231). Semi-quantitative RT-PCRs for the estimation of expression of growth factors and cytokines were performed in cultured breast cancer cells in various concentration of RANKL. For invasion analysis, we used matrigel invasion assay. Enzymatic activities of MMP-2, 9 were identified using zymography.
Results
We have shown the NF-B activation in MCF-7 & MDA-MB-231 cells when they were incubated with soluble RANKL (1~100 ng/ml). Semi-quantitative RT-PCR showed that the expressions of PTHrP, TGF mRNA in ratio to internal standard GAPDH mRNA were elevated in RANKL-treated cells compared with control (respectively; p<0.05), but the levels of IL-1, IL-6, OPG mRNA were not changed. MDA-MB-231 cells supplemented with RANKL for 18 hr in 0.1% BSA/-MEM dose- dependently showed the high invasiveness (control: 16±5 vs 100 ng/ml of RNAKL: 146±27). Zymography and RT-PCR analysis of MMP-2, MMP-9 which were correlated with cancer invasion were not changed.
Conclusion
These results suggest that the locally increased RANKL in bone microenvironment may activate the invasiveness of breast cancers and enhance expression of osteolytic factors in the tumor cells.
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| Key Words:
Breast cancer, RANKL, PTHrP, TGF |
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