Expression of transduced HSP70 gene protects chondrocytes from stress

Objective. To investigate thr efficacy of adenovirus vector mediated transd uction of heat shock protein 70 (HSP70) gene to human chondrocyte-like cell (HCS-2/8) against heat stress. Methods. Two adenovirus vectors that contain wild-type (AxSHEwt) or mutant- type (AxSHEmt) HSP70 gene, and that are regulated by SR alpha promoter, wer e constructed. The mutant-type lacks the area that expresses stress durabil ity. One of the 2 adenovirus vectors was added to the cultures of human cho ndrocyte-like cells (HCS-2/8). Heat stress (48 degreesC) was applied to the transduced cells for 2 h. and the efficacy of adenovirus vector mediated t ransduction of HSP70 gene against heat stress in the chondrocytes was inves tigated using alamar blue assay and MTT assay. Results. Absorbance levels at 48 degreesC were 300.3 +/- 51.9 and 1.173 +/- 0.011 in the controls, 278.5 +/- 33.8 and 1.217 +/- 0.018 in the AxSHEmt t ransduced cells, and 349 +/- 14.7 and 1.371 +/- 0.033 in the AxSHEwt transd uced cells. The level in the AxSHEwt transduced cells was significantly hig her than in the other 2 groups (p < 0.05). With 37<degrees>C treatment, no significant difference was observed. Conclusion. Chondrocytes to which HSP70 gene was transduced had a significa ntly higher metabolic activity and viability under heat stress.