Introduction. Heme oxygenase-1 (HO-1) was shown to regulate proliferation in a cell type specific manner. It inhibits growth of epithelial and smooth muscle cells, but increases proliferation of endothelium and many cancer cells. It is also well known that HO-1 protects both normal and cancer cells under oxidative stress conditions. The aim of our study was to investigate the role of HO-1 in proliferation and cell cycle control of murine primary fibroblasts. Moreover we determined the effect of HO-1 in protection of these cells against oxidative stress and cytostatics.

Methods and results. Experiments were performed on primary fibroblast isolated from HO-1-/-, HO-1+/- and HO-1+/+ mice. Proliferation and migration were stimulated by increasing the concentration of fetal bovine serum (FBS) up to 10% after a 24 h starvation of cells in medium containing 0.2% FBS. We found that the lack or low expression of HO-1 (HO-1-/- and HO-1+/-, respectively) significantly increased serum-induced proliferation of fibroblasts when compared to the wild type cells. This was caused by increased proportion of cells at the S-phase. In contrast, migration of HO-1-/- fibroblasts was weaker than that of HO-1+/- and much weaker than HO-1+/+. Although preliminary screening using GEArray membranes suggested HO-1 dependent differences in expressions of several genes involved in the cell cycle regulation, the exact analyses done by real-time PCR, showed no changes in the genes tested, namely in cyclin D1, cyclin D3, p21 or cyclin dependent kinase 4. On the other hand, we demonstrated that fibroblasts isolated from HO-1-/- mice were the most prone to death caused by hemin, which corresponds to previous reports and proves proper functioning of the model. Based on the quantitative PCR analysis of genes involved in cellular protection against oxidative stress we found that expression of super oxide dismutase-2, ferritin and glutathione transferase was significantly higher in HO-1+/+ than in HO-1-/- fibroblasts exposed to hemin, whereas there were no differences in the expression of super oxide dismutase-1, glutathione reductase, tioredoxin and catalase. Surprisingly, cells genotype did not influence survival of cells after exposure to hydrogen peroxide or doxorubicin. Moreover, HO-1+/+ cells were even less protected from toxic effect of cisplatin than HO-1-/-.

Conclusion. Expression of HO-1 decreases proliferation and increases migration of primary fibroblasts. Cytoprotective effects of HO-1 are evident in cells treated with hemin, but not in cells exposed to doxorubicin, cisplatin or hydrogen peroxide.

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