Molecular Pathology of Highly Metastatic Rat Osteosarcoma

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1) We applied wild-type p53 gene transfer to the rat osteosarcoma cells by lipofection, and reconstitution of the p53 inhibits cellular growth in rat osteosarcoma COS1NR cell line, and this growth-suppressive effect is partly due to apoptosis involving bcl-2 gene suppression.2) We investigated the p16 and p19gene expression in rat osteosarcoma cell lines, COS1NR, COS2NR and COS4NR and their subcutaneous tumors and lung metastatic nodules, and rat malignant fibrous histiocytoma cell lines, MFH1NR, MFH2NR and their subcutaneous tumors by RT-PCR.The results showed that there were no differences of the expression of p16 and p19 genes among the all cell lines and tumors. The PCR-SSCP analysis also showed no deletions and point mutations in any genes in any tumors investigated.3) We investigated TGF-β isoforms by Multi-probe RNA protection assay in COS1NR, COS2NR, COS4NR, MFH1NR and MFH2NR.The results showed TGF-β 1to be strongly expressed and no TGF-β 2 and 3 detected in MFHs, while TGF-β 1 and 2 was found to be moderately and 3 weakly expressed in OS cell lines. No changes in TGF-β type II receptor expression, and no mutations of TGF-β type II receptor, Smad2 and 4 genes were detected by RT-PCR and RT-PCRrestricted SSCP anaysis.