Serçin Özlem Çalişkan

This study aimed to characterize the chemical constituents of the methanolic extract of Euphorbia anacampseros using high‐performance liquid chromatography (HPLC–DAD) and gas chromatography–mass spectrometry (GC–MS) analyses. The cytotoxic activity of the extract was assessed using the MTT assay, and its light‐enhanced cytotoxic effects were evaluated to investigate its potential for future photodynamic applications. Additionally, molecular docking analysis was performed to evaluate interactions between the identified compounds and the porphobilinogen deaminase (PBGD) enzyme. The mean inhibitory concentration (IC ₅₀ ) values were 54.72 µg/mL for HT‐29, 50.34 µg/mL for MCF‐7, and 58.97 µg/mL for A549 cell lines. In the dark, the extract exhibited moderate and nonselective baseline cytotoxicity across all tested cell lines, with IC ₅₀ values ranging from 46.95 to 58.97 µg/mL. However, the cytotoxic activity of the extract in HT‐29 cells was significantly enhanced following irradiation with blue, green, yellow, or red light, suggesting its potential for future photodynamic applications. Molecular docking targeting PBGD revealed that rutin and epicatechin exhibited the highest binding affinities, with binding energies of –9.9 and –7.8 kcal/mol, respectively. These findings suggest that E. anacampseros constituents may act as photosensitizing agents, potentially enhancing photodynamic therapy through interactions with enzymes involved in porphyrin metabolism.

Rationale: In this study, the phytochemical composition, in vitro anticancer activity, photodynamic therapy (PDT) activity, and molecular interaction potential with carbonic anhydrase IX (CA IX) of Cakile maritima methanol extract were evaluated. Methods and Results: LC–MS/MS analysis identified 11 compounds among 35 screened phytochemicals, with fumaric acid (13797.19 ppb), ferulic acid (771.37 ppb) and p‐coumaric acid (601.95 ppb) accounting for approximately 97% of the total content. Molecular docking results demonstrated that sinapic acid (−6.13 kcal/mol) and quinic acid (−6.11 kcal/mol) exhibited the highest binding affinities toward CA IX. Cell culture studies revealed that LogIC ₅₀ values differed significantly among cell lines ( p < 0.0001). PDT applied to the MKN‐28 gastric cancer cell line significantly enhanced cytotoxicity compared with the control group, with the highest photodynamic efficacy observed under green‐light irradiation ( p < 0.0001). Conclusions: These findings indicate that C. maritima methanol extract exhibits light‐enhanced cytotoxicity in MKN‐28 gastric cancer cells and warrant further investigation regarding its potential photosensitising properties.

Objectives: In the treatment of acute myeloid leukemia (AML), conventional therapies can lead to severe side effects and drug resistance. There is a need for alternative treatments that do not cause treatment resistance and have minimal or no side effects. Sonodynamic therapy (SDT), due to its noninvasive, multiple repeatability, localized treatment feature and do not cause treatment resistance, emerges as an alternative treatment option. However, it has not received sufficient attention in the treatment of AML especially acute promyelocytic leukemia (APL). The aim of the study was to investigate the potential differentiation and antileukemic effects of acridine orange (AO)‐mediated SDT on HL60 cells.Methods: Cell viability was determined by the 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide (MTT) method in the control, ultrasound, AO concentrations, and ultrasound‐exposed AO concentrations groups. Transmission electron microscopy (TEM) was used to determine morphology, and flow cytometry was used to determine apoptosis, DNA cycle, cell volume, mitochondria membrane potential (Δψm), reactive oxygen species (ROS) production, and differentiation markers (CD11b and CD15) expressions. Additionally, toluidine blue staining for semithin sections was used to determine differentiation.Results: The cytotoxicity of AO‐mediated SDT on HL60 cells was significantly higher than other groups, and TEM images showed that it caused various morphological changes typical for apoptosis. Flow cytometry results showed the presence of early apoptosis, subG1 arrest, loss of Δψm, increase of intracellular ROS production, decreased cell volume, and increased expression of CD11b (1.3‐fold) antigen and CD15 (1.2‐fold) antigen.Conclusion: Data showed that AO‐mediated SDT significantly induced apoptosis in HL60 cells. Increased expression of CD11b and CD15 antigens and morphological findings demonstrated that AO‐mediated SDT contributes to granulocytic differentiation in HL60 cells. AO‐mediated SDT has potential as an alternative treatment of APL.

Aim: Sonodynamic antitumor therapy is a promising, novel method for the treatment of cancer. To determine the effects of malachite green (MG) in the presence of ultrasound (US), MG was tested in vitro on HL60 cells at different concentrations as a sonodynamic compound. We investigated cell viability, morphology, and the occurrence of ER stress after MG-mediated sonodynamic therapy (SDT) in HL60 cells. Aim: Sonodynamic antitumor therapy is a promising, novel method for the treatment of cancer. To determine the effects of malachite green (MG) in the presence of ultrasound (US), MG was tested in vitro on HL60 cells at different concentrations as a sonodynamic compound. We investigated cell viability, morphology, and the occurrence of endoplasmic reticulum (ER) stress after MG-mediated sonodynamic therapy (SDT) in HL60 cells. Material and Method: Four groups were formed, including a control group, a group subjected to ultrasound (US) only, a group treated with various concentrations of MG, and a group treated with US using the same concentrations. The cells were treated with 1MHz ultrasound at 2 W/cm2 for 3 minutes. The assessment of cell viability was conducted 24 hours post-treatment through the utilization of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cell morphology and apoptotic index were determined using Giemsa staining, while GRP78 and PERK expressions were determined through immunocytochemistry staining. Results: The cell cytotoxicity of HL60 cells significantly increased after MG-mediated sonodynamic therapy. After treatment, apoptotic cells with micronuclei were observed morphologically. Significant levels of GRP78 and PERK expression were observed in all groups, except for PERK expression in the US group, compared to the control group. Conclusion: The induction of ER stress, accompanied by intense apoptosis and a marked decrease in cell viability, demonstrates the potential of MG-mediated sonodynamic therapy in cancer treatment. Investigating ER stress as a molecular target may contribute to improving the treatment method.

Aim: Our objective is to explore the relationship between the anti-leukemic impact of malachite green-mediated photodynamic therapy (PDT) and the induction of endoplasmic reticulum (ER) stress in acute promyelocytic leukemia cells (HL-60). Material and Method: For one hour the cells were incubated with different concentrations (3.125, 1.56, 0.78, 0.39, 0.195, 0.0975, 0.04875 μM) of malachite green and then were exposed to 0.47 mW/cm2 irradiance and 0.84 J/cm2 fluence for 30 minutes. Also, HL-60 cells were exposed to PDT with light only and both in the presence or absence of malachite green. MTT assay was used to determine cell viability, and immunocytochemical staining was used to detect the expression of ER stress markers Protein Kinase R-like ER Kinase (PERK) and Glucose-regulated protein 78 (GRP78). Results: The cell viability of the treatment group (combination of malachite green and light) was significantly decreased compared to the malachite green, control group, and light control. Moreover, immunocytochemical staining scores showed that PERK and GRP78 were significantly upregulated in the treatment group compared with other groups. Conclusion: Our results indicate that ER stress may contribute to the cytotoxicity occurring in HL-60 cancer cells after malachite green-mediated PDT. Future studies will be crucial in shedding light on the molecular mechanisms underlying ER stress that may occur after PDT. These findings lay the foundation for further investigations in this area.

Background/aim: Photodynamic therapy (PDT) has received great attention over the past decade in the treatment of diseases such as leukemia which is a cancer of the blood and bone marrow cells that causes a significant number of deaths worldwide. In this study, it was aimed to investigate the effects of Nile blue-mediated PDT (NB-mediated PDT) on HL60 cells. Materials and methods: The effect of NB-mediated PDT on cell proliferation was evaluated with cell volume analysis using flow cytometry at 24 h. Cell apoptosis, ROS production, mitochondrial membrane potential, and cell cycle analysis were evaluated using annexin V-FITC, H2 DCFDA, JC-1, and PI staining, respectively, by flow cytometry and fluorescence microscopy. The morphological and ultrastructural analyses were examined by Giemsa staining and SEM. CD11b staining is used to determine the differentiation of leukemia cells. Results: NB-mediated PDT induced an apoptotic response at 12.5 µM in HL60 cells. When Giemsa staining and SEM images were evaluated, apoptotic bodies, holes, and occasional folds were detected on the surfaces of cells in the NB-mediated PDT group. Conclusion: The NB-mediated PDT had no effect on the differentiation of leukemia cells, but this therapy affects the growth of HL60 cells in vitro, which may provide a new idea for removing leukemic cells from bone marrow intended for autologous transplant.

Aim: Acute myeloid leukemia (AML) is a disease characterized by relapse and treatment resistance in most patients. Therefore, there is a need for targeted therapies in AML. Photodynamic therapy (PDT) is a promising alternative for the treatment of malignant tumors. Also, PDT has the potential to be used individually or complementally in the treatment of leukemia. In this study, it was aimed to investigate possible the effect of malachite green (MG)-based PDT on acute myeloid leukemia cells. Materials and Methods: Cells were incubated with 0.19, 0.39, 0.78,1.56, 3.125, and 6.25 µM MG for one hour and irradiated with 46.4 J/cm2 of light. The trypan blue test was used to assess the viability of cells, and the change in mitochondrial activity was determined by MTT. Morphological features were determined by Giemsa staining and scanning electron microscopy. Cell cycle and Annexin V/PI assays (measuring fluorescence emitted by staining reagents) were measured by flow cytometry. Results: With the combination of MG and light, HL60 cell viability was found to be significantly reduced compared to the control group. Giemsa staining and SEM results showed that 3.125 μM MG-based PDT induced various morphological changes in cells typical for apoptosis. Late apoptosis was observed in cells treated with 3.125 μM MG combined PDT according to Annexin/PI staining, further showing that it caused an arrest in the subG1 phase of the cell cycle. Conclusion: MG-based PDT has the potential to inactivate HL60 cells. Thus, MG-based PDT may ensure a promising approach for treating acute myeloid leukemia cells.

OBJECTİVE: In this study, we aimed to examine of antileishmanial effect of curcumin-mediated sonodynamic (SDT), photodynamic (PDT), and sonophotodynamic (SPDT) therapies with a potential mechanism against the Leishmania tropica (L.tropica) promastigotes. SPDT is a new treatment modality for Leishmaniasis that combines photodynamic and sonodynamic therapies. Curcumin is a natural antiinflammatory agent that has been used for treating medical conditions for many years. Curcumin was used in this study both as a sonosensitizer and photosensitizer to compare the effect of PDT, SDT, SPDT on L.tropica promastigotes. MATERIALS AND METHODS: The cells were incubated with different concentrations (0.25, 1,4,16 and 64μM) of curcumin for 1 hour, were exposed to 3 W/cm2 intensity ultrasound for 1MHz frequency and/or subjected to 1,32 J/cm2 light irradiation for 30 minutes. Also, parasite cells were exposed for SPDT with ultrasound and light only and both in the presence or absence of curcumin. XTT was used to evaluate cell viability and giemsa staining was used to determine morphological changes. RESULTS: With the combination of curcumin and ultrasound, curcumin and light, curcumin mediated ultrasound and light,L.tropica promastigote viability was found to be decreased compared to the control, ultrasound-control and light-control group. The greatest reduction was found in the SPDT group. Giemsa staining findings showed that curcumin-mediated SDT, PDT and SPDT induced several morphological alterations in L.tropica promastigotes atypical. These results showed that SPDT is more effective than other therapies on L.tropica promastigotes CONCLUSIONS: Curcumin-mediated SPDT may provide a promising approach for L.tropica promastigotes.