DNA damage and antioxidant properties of CORM-2 in normal and cancer cells

Abstract

In this study, we compared the effect of tricarbonyldichlororuthenium (II) dimer (CORM-2) and its CO-depleted molecule (iCORM-2) on human peripheral blood mononuclear cells (PBMCs) and human promyelocytic leukemia HL-60 cells. We determined cell viability, DNA damage and DNA repair kinetics. We also studied the effect of both compounds on DNA oxidative damage, free radical level and HO-1 gene expression. We showed that at low concentrations both CORM-2 and iCORM-2 stimulate PBMCs viability. After 24-h incubation, CORM-2 and iCORM-2, at the concentration of 100 µM, reduce the viability of both PBMCs and HL-60 cells. We also demonstrated that CORM-2 and iCORM-2, in the 0.01-100 µM concentration range, cause DNA damage such as strand breaks and alkaline labile sites. DNA damage was repaired efficiently only in HL-60 cells. CORM-2 significantly reduces oxidative stress induced by 1 mM H 2 o 2 in normal and cancer cells. On the contrary, iCORM-2 in HL-60 cells increases the level of free radicals in the presence of 1 and 5 mM H 2 o 2. We also revealed that both CORM-2 and iCORM-2 induce HO-1 gene expression. However, CORM-2 induces this gene to a greater extent than iCORM-2, especially in HL-60 cells at 100 µM. Finally, we showed that CORM-2 and iCORM-2 reduce H 2 o 2-induced DNA oxidative damage. Furthermore, CORM-2 proved to be a compound with stronger antioxidant properties than iCORM-2. Our results suggest that both active CORM-2 and inactive iCORM-2 exert biological effects such as cyto-and genotoxicity, antioxidant properties and the ability to induce the HO-1 gene. The released CO as well as iCORM-2 can be responsible for these effects. Carbon monoxide (CO) is a colorless, tasteless and odorless gas produced by the burning of fuels and organic materials. It is reported to be the most frequent cause of fatal poisoning with an incidence rate of 31%. CO is readily absorbed and is unchanged by the lungs. CO demonstrates more than 200-fold stronger affinity for hemoglobin compared to oxygen. Therefore, even a small level of CO may cause poisoning. In contrast to hypoxiainducing toxic concentrations, a low dose of CO or even nanomolar concentrations exert biological activities. CO is produced in low amounts as a byproduct of normal human metabolism by the enzyme called heme oxygenase (HO-1) 1. CO has the ability to reduce the stimulation of guanylate cyclase to generate cyclic guanosine 3′,5′-monophosphate (cGMP). As a signaling molecule, CO modulates several p38 mitogen-activated protein kinase (MAPK)-related signaling pathways via both cGMP-dependent and independent processes, directly activates calcium-dependent potassium channels and induces protein kinase B (Akt) phosphorylation via the phosphatidylinositol 3-kinase/Akt pathway 2. Moreover, CO inhibits mitochondrial respiration by binding the ferrous heme a 3 in the active site of cyclooxygenase (COX), effectively shutting down oxidative phosphorylation, similar to the effects of cyanide and nitric oxide (NO) 3. The cGMP-dependent activity of CO includes inhibit...

Kaempferol and Its Glycoside Derivatives as Modulators of Etoposide Activity in HL-60 Cells

“…The HL-60 cells were seeded on 96-well plates at a density of 1 × 10 4 cells/mL in a culture medium and incubated for 24 h with the tested compounds (etoposide, kaempferol, P2, P5, P7) at 37 °C in 5% CO 2 . The assay was described in detail by Juszczak et al (2020) [ 44 ].…”

Kaempferol and Its Glycoside Derivatives as Modulators of Etoposide Activity in HL-60 Cells

“…The HL-60 cells were seeded on 96-well plates at density of 1 × 10 4 cells/ml in a culture medium and incubated for 24 h with the tested compounds (etoposide, kaempferol, P2, P5, P7) at 37C in 5% CO2. The assay was described in detail by Juszczak et al (2020) 9 .…”

“…The cells incubated for 15 min with 5 mM H2O2 at 37°C were the positive control. The assay was described in detail by Juszczak et al (2020) 9 .…”

“…(EurX, Gdansk, Poland), according to the manufacturer's instructions. The assay was described in detail by Juszczak et al (2020) 9 .…”

A red light‐activable MnI(CO)3‐functionalized gold nanocomposite as the anticancer prodrug with theranostic potential

“…However, the hepatotoxicity and prolonged photosensitivity of the porphyrin-based PDT agent and the lack of an ideal photosensitizer have halted the clinical use of PDT. [9][10][11][12][13][14] Several photoactivable transition metal-based compounds and nanohybrids that generate reactive oxygen species under visible-light activation have emerged as viable alternatives to porphyrin-based photochemotherapeutic agents. [15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] The salutary effect of a low dose of CO has prompted research on the photoactivated release of exogenous CO from CO-releasing molecules (CORMs) with spatial and temporal resolution for clinical applications, including cancer therapy.…”