To study the ef fect of extracellular calcium on metastatic behavior, we quantified the chemotactical migration and cell prolifer ation of these RCC cells under calcium influence. The molecular mechanisms accountable for the effects ob served were analyzed by quantifying the activity of intra cellular signaling pathways, especially the AKT and MAPK pathways and its regulatory phosphatase PTEN. The elucidation in the importance of calcium and CaSR in the process of bone metastasis could reveal new prog nostic markers and contribute towards the development of new target therapies. Benefits Tissue specimens of RCC sufferers creating bone metastases show a high CaSR expression Quantification on the CaSR expression in RCC was per formed by analyzing tumor and regular tissue specimens from RCC patients without having metastases and from individuals creating lung or bone metastases within 5 years after nephrectomy by quantitative RT PCR.
The outcomes have been correlated with the localization on the metastatic web pages. In tumor specimens of individuals devel oping bone metastases, CaSR mRNA expression was 7. 9 fold larger than in tumor specimens of individuals with no metastases. Tumor specimens from individuals with no metastases or with lung metastases expressed read this post here CaSR mRNA moderately. In regular renal tissue, CaSR ex pression was considerably greater than in tumor speci mens. In regular renal tissue of sufferers building bone metastases, CaSR mRNA expression was 1. eight fold larger than in specimens of sufferers without metastases. Analyzing the CaSR protein within the tissue specimens we observed a comparable trend, while the impact was even less pronounced.
Bone metastatic major RCC cells show a high CaSR expression The expression of CaSR in principal RCC cells was deter mined by flow cytometry. Corresponding for the final results obtained from tissue specimens, CaSR selleck chemicals expression in RCC cells cultivated from sufferers developing bone me tastases was 3. 7 fold higher than in cells from sufferers with no metastases. In cells from individuals de veloping lung metastases, CaSR expression was 1. 9 fold greater than in non metastasizing RCC cells. Therapy with 5 mM calcium had no influence on CaSR expres sion of RCC cells. Extracellular calcium stimulates migration and proliferation of bone metastasizing main RCC cells Since the CaSR expression was enhanced in tumor tissue and key cells from individuals who developed bone me tastases, we investigated the influence of extracellular cal cium in processes of metastasis.
The migratory possible of major RCC cells was analyzed inside a Boyden chemo taxis chamber making use of calcium as chemotaxin. To investi gate the influence of calcium on proliferation of those major RCC cells, they were incubated with calcium for 30 min and cell proliferation was determined by BrdU in corporation.

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