Mutations of the p53 tumor suppressor gene in rheumatoid arthritis synovium / 대한정형외과학회잡지

PURPOSE: To analyse the expression of the p53 tumour suppressor gene in the synovial tissue from rheumatoid patients. Material and Methods : Synovial membranes were obtained from 13 patients diagnosed as having RA, and 9 osteoarthritis (OA) patients. We studied p53 expression by immunohistochemical analysis and p53 DNA sequence using direct DNA preparation method. RESULT: In immunohistological studies, the Do-1 monoclonal antibody stained at 6 specimens out of the 13 rheumatoid arthritis tissue biopsies analysed. There was no p53 mutation in osteoarthritis samples, but there were 4 p53 mutations from the 13 rheumatoid arthritis samples. p53 mutations were found at the codon 177 (CTG to CTA, GA), 277 (TGT to TGC, TC), and, two patients at the codon 237 (CAT to TAT, CT). CONCLUSION: The predicted amino acid substitutions in p53 were similar to those commonly observed in a variety of tumors and might influence growth and survival of rheumatoid synoviocytes.

Gene Transfer Into Human Chondrocyte Derived Cells Using A Liposome Mediated Transfection System / 대한정형외과학회잡지

PURPOSE: To introduce the CMV promoter driven luciferase and -galactosidase marker gene into previously permeabilized human chondrial cells. MATERIALS AND METHODS: The cultured chondrocyte cells were transfected with a liposome/DNA mixture (pCMV-Luc or pSV40-lacZ). Cultured cells not transfected by liposome/DNA were used as a control. After forty-eight hours of incubation, the cells were used for reporter gene assays and the polymerase chain reaction (PCR). RESULTS: Fifteen percent of the chondrocyte cells treated with liposome/ pSV40-lacZ DNA were positive for -gal staining. Chondrocyte cells transfected with pCMV-Luc yielded a 70-fold increase in luciferase activity over that of the control cells. A PCR product corresponding to the luciferase gene appeared only in the transfected chondrocyte cells. These results indicate that the human chondrocyte cells can be transfected with pCMV-Luc and pSV40-lacZ. CONCLUSION: This system is particularly suitable for gene therapy, as well as for the use of genetically modified cartilage cells for resurfacing full thickness articular cartilage defects.