Experimental Research On The TIMP-3 Gene Promoter Methylation And Protein Expression In Joint Cartilage Of Osteoarthritis Patients

Objectives To investigate the role of Tissue inhibitor of metalloproteinase-3 (TIMP3) gene promoter methylation in the occurrence and development of osteoarthritis (OA).Methods General DNA were dealed with Sodium Bisulfite method. The methylation of TIMP3 promoter was detected by Methylation-specific PCR (MSP). TIMP3 protein expression in joint cartilage cells were detected by Immunohistochemical method.35 OA patients and 14 control patients were recruited from the in-patient of the second Xiangya hospital of Central South University.The results were analyzed byχ2 or Fisher's test and Spearman correlation analysis of SPSS 17.0 statistic analysis software.Results1. The positive percent of TIMP3 promoter methylation in OA patients and healthy persons were 74.3%(26/35) and 35.7%(5/14) respectively, Usingχ2 test, there was significantly different between the two groups(χ2=6.402,P=0.0114).2. The positive percent of TIMP3 protein in OA patients and healthy persons were31.4%(11/35) and 71.4%(10/14) respectively, Usingχ2 test, the positive percent of TIMP3 protein in OA patients was significantly lower than that in healthy persons(χ2=6.533,P=.0106).3. Prmoter methylation and protein expression level of TIMP3 gene showed significantly negative correlation.ConclusionsTIMP3 promoter is hypermethlated in joint cartilage cells of OA patients. The hypermethylation affect the protein expression of TIMP3, which might take a role in the occurrence and development of OA...