University of Health and Sciences, Istanbul Training and Research Hospital, Department of General Surgery - Istanbul, Turkey.
Istanbul Health and Technology University, Faculty of Pharmacy, Department of Biochemistry - Istanbul, Turkey.
Rev Assoc Med Bras (1992). 2022 Apr;68(4):507-513. doi: 10.1590/1806-9282.20211193.
We aimed to examine the potential anticancer effects of ozone applied after chemotherapeutic treatment with different concentrations of doxorubicin in Luminal-A subtype of human breast cancer cell line (MCF-7) and compare the results with effects on L929 fibroblast cell line.
Both cell lines were incubated with increasing doses of doxorubicin (1-50 μM) for 24 h at 37°C. Then, half of groups were incubated with 30 μg/mL ozone for 25 min as combination groups. Cell viability was analyzed by MTT assay, apoptosis by flow cytometry, and levels of tumor necrosis factor alpha, transforming growth factor beta, and matrix metalloproteinase-2 and MMP-9 by immunocytochemistry.
Doxorubicin + ozone treatment enhanced viability of L929 (p<0.01) but reduced viability of MCF-7 compared to only doxorubicin-applied cells without ozone treatment (p<0.001). This combined treatment also enhanced apoptotic effect of doxorubicin on MCF-cells (p<0.001), but not on L929. It significantly increased all protein levels of L929 compared with those of other groups (p<0.05 for tumor necrosis factor alpha and MMP-2; p<0.01 for transforming growth factor beta and MMP-9). This treatment reversed the effect of doxorubicin on tumor necrosis factor alpha levels and considerably reduced MMP-2 and MMP-9 levels of MCF-7 compared with those of control group (p<0.01 and p<0.001, respectively).
Ozone treatment potentiated the apoptotic and anticancer activities of doxorubicin in MCF-7 cells and showed repairing and healing effect on healthy fibroblast cells, which were damaged from cytotoxic effects of chemotherapeutic agent. MCF-7 cells may acquire sensitivity against the doxorubicin combined with ozone treatment through activating tumor necrosis factor alpha, MMP-2, and MMP-9 expressions.
本研究旨在探讨不同浓度多柔比星化疗后应用臭氧对人乳腺癌 Luminal-A 亚型细胞（MCF-7）的潜在抗癌作用，并与对 L929 成纤维细胞的作用进行比较。
将两种细胞系分别在 37°C 下孵育不同剂量的多柔比星（1-50 μM）24 小时。然后，将一半的细胞组与 30 μg/mL 的臭氧孵育 25 分钟，作为联合组。通过 MTT 分析检测细胞活力，通过流式细胞术检测细胞凋亡，通过免疫细胞化学检测肿瘤坏死因子-α、转化生长因子-β以及基质金属蛋白酶-2 和 MMP-9 的水平。
与未用臭氧处理的多柔比星处理组相比，多柔比星+臭氧处理组增强了 L929 细胞的活力（p<0.01），但降低了 MCF-7 细胞的活力（p<0.001）。与其他组相比，这种联合处理还增强了多柔比星对 MCF-7 细胞的凋亡作用（p<0.001），但对 L929 细胞无此作用。与其他组相比，它显著增加了 L929 细胞的所有蛋白水平（肿瘤坏死因子-α和 MMP-2 为 p<0.05；转化生长因子-β和 MMP-9 为 p<0.01）。与对照组相比，这种处理逆转了多柔比星对肿瘤坏死因子-α水平的作用，并显著降低了 MCF-7 细胞中 MMP-2 和 MMP-9 的水平（分别为 p<0.01 和 p<0.001）。
臭氧处理增强了 MCF-7 细胞中多柔比星的凋亡和抗癌活性，并对因化疗药物细胞毒性而受损的健康成纤维细胞显示出修复和愈合作用。MCF-7 细胞可能通过激活肿瘤坏死因子-α、MMP-2 和 MMP-9 的表达对多柔比星联合臭氧治疗产生敏感性。
