Selection Of Metastatic And Non-metastatic Cells From Human Primary Renal Cell Carcinoma (RCC) In Vitro And In Vivo

[Objective]To select metastatic or non-metastatic RCC from the same sources and discriminate the biological characteristics of the two for the purpose of looking for the biomarker of metastatic RCC.[Methods]1. Fresh RCC surgical specimens were obtained from 32 primary RCC patients. Tumor specimens were cultured following enzyme digestion or minimized fragment in vitro. Morphological characteristics and doubling time of the cells were subsequently determined. Karyotypic analysis and anchorage-independent growth were then evaluated.2. Invasion assay in vitro using the Transwell cultures were performed for the selection of highly invasive potential cells of primary renal cancer cells.3. Surgical specimens/tumor cell suspensions were transplanted/injected into the various tissues of BALB/c nude mice (subcutis, perinephrium, COl (cellular orthotopic injection into renal capsule), SOI (surgical orthotopic implantation into renal capsule)). Tumorigenicity and metastasis were observed. Expression of VEGF, bFGF, P16, bcl-2 and c-met in primary and metastatic RCC from the same nude mice was determined, respectively.[Results]1. The success rate of primary culture was as high as 90.6% (29/32), while long-term cultures were difficult. The renal cancer cells expressed heterogeneous shape and size, and the some were fibroblast-like or growing in swirls that were very similar to normal kidney epithelial cells. The ranges of doubling time were from 14.5h to 101h. The ten percent of cells formed clones in the soft agar. The mode of chromosome was 48, indicating that abnormal...