| Prostate cancer is a common malignant tumor in male reproductive system, the morbility of which showed an increasing trend with the improvement of people’s living standards. Studies have shown that high-fat western diet may be associated with the occurrence and development of prostate cancer. Although many studies have explained the influence mechanisms of PUFAs on cancer, but it is still a lack of research on the biological mechanisms of fatty acids in cancer cells and normal cells. In this study, a preliminary study was done to investigate the impact of PUFAs in fish oil on prostate cells (PC-3and RWPE-1). And then representative n-3PUFAs (ALA, EPA, DHA) and n-6PUFAs (LA, GLA, AA) were selected to study their effect on cellular antioxidant system, metal ions absorption and inflammatory responses in two cell lines, and the metabolic transformation of these PUFAs in cells.MTT results showed that fish oil is almost harmless for RWPE-1, but can partly kill PC-3. Apoptosis analysis by flow cytometry showed that PC-3had higher rate of apoptotic cells than RWPE-1when treated with the same dose of fish oil. Fish oil damaged the cellular antioxidant systems of two cell lines and RWPE-1had higher MDA accumulation than PC-3through SOD, MDA, CAT and GSH-Px analyses. GC results showed that there were significant differences of fatty acid compositions and proportions of different kinds of fatty acids in two cell lines, and GLA, AA, EPA and DHA contents in PC-3had greater increase than RWPE-1supplemented with fish oil. Inflammatory cytokines were also determined by ELISA, and the results showed that LXA4secretion increased significantly in PC-3,which almost remained at the same level in RWPE-1, and the expression of TNF-a and IL-6were significantly decreased in PC-3, which were gradually increased in RWPE-1.Studies of cellular injury induced by PUFAs showed that PUFAs inhibited the growth of both cell lines, in lower damage with LA and ALA, but in serious injury with GLA and AA. AA, GLA, EPA and DHA had larger influences on membrane permeability and mitochondrial membrane potential in both cell lines than LA and ALA, and also had greater effect on PC-3. HPLC results revealed that PUFAs can inhibit the synthesis of ATP in PC-3, wherein AA, GLA, EPA and DHA showed greater inhibition than LA and ALA, while ATP levels in RWPE-1cells changed little.The influences of PUFAs on cellular antioxidant systems of two cell lines were investigated, and the results indicated that ROS accumulation in RWPE-1was significantly higher than PC-3. AA and GLA significantly reduced GSH-Px activity in RWPE-1, and GSH-Px activity in PC-3was significantly decreased with EPA and DHA. GSH levels increased in two cell lines treated with PUFAs. PUFAs had less influences on SOD activity and NO content, but increased MDA accumulation in two cell lines especially in supplementation with, AA, GLA, ALA and DHA.We investigated the effects of PUFAs on inflammatory responses and their biotransformation in cells, and found that PC-3had more obvious lipid droplets than RWPE-1, and lipid droplets in n-6PUFAs groups were more intensive than that in n-3PUFAs groups by oil red staining. Supplementation with PUFAs increased the contents of specific fatty acids and changed fatty acid profiles in cells significantly by GC analysis. ELISA detection also showed that there were significant changes in IL-6, TNF-a and LXA4expression by two cell lines treated with PUFAs.We studied the impact of PUFAs on Zn2+, Cu2+, Cd2+absorption by cells, and results showed that PC-3had better tolerance to Zn2+at low concentrations than RWPE-1, but it was opposite at high concentrations. Both two cell lines had high tolerance to Cu2+, while RWPE-1suffered heavier damage with prolonged treatment. Cd2+induced significant cytotoxicity on two cell lines, but was more harmful to PC-3. AAS analysis concluded that PUFAs had less impact on Zn2+uptake with two cell lines, but can inhibit Cd2+absorption and promoted Cu2+uptake to some extent. |
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