impact-different-extraction-methods-on-vitro-bioactivity-profiles

Impact of Different Extraction Methods on the <em>In Vitro</em> Bioactivity Profiles of <em>Terfezia Claveryi</em>

Terfezia claveryi is an edible desert truffle traditionally consumed in arid regions. In recent years, it has attracted increasing scientific interest due ...

Kirsehir Ahi Evran University

Low-dose chemotherapy in advanced stages of cancer does not give a positive response in treatment. The use of high-dose antineoplastic drugs creates significant side effects. The limiting situation in treatment creates a need for new generation drugs with less side effects and new treatment methods that will enable low-dose drug use. Electroporation (EP), a phenomenon, is a technique in which the membrane permeability is increased by the establishment of hydrophilic pores in the cell membrane with short and high-voltage electrical pulses. In the present investigation study, we aimed to inspect the effects of EP plus bleomycin, cisplatin, and calcium administration (CaEP) on cell viability, apoptotic activity, gene expression p53, Bax/Bcl-2 rate mitochondrial membrane potential (ΔΨm), and cell cycle in HeLa cervical cancer cell line. The permeabilization of the membrane was evaluated in flow cytometry with the PI method, and cell viability was measured in an ELISA reader with the WST-8 method. For bleomycin and cisplatin doses applied to HeLa cells, the concentration values (IC) that inhibited 50% of the cells were found to be 214.11 ± 4.7 µM and 35.16 ± 3.3 µM, respectively. The IC values of the groups administered together with EP were calculated as 0.44 ± 0.3 µM for bleomycin and 20.55 ± 4.3 µM for cisplatin. There was no change in cell viability in calcium alone application, but a statistically notable reduction in cell vitality was observed in CaEP application. An increase in ΔΨm was found in bleomycin and CaEP exposure with EP. It was determined that EP exposure caused G0/G1 arrest in the cell cycle at all electric field intensities. It was determined that EP application in HeLa cells increased bleomycin cytotoxicity 487 times and cisplatin cytotoxicity 1.71 times, and CaEP could be an alternative treatment method.

Treatment of cervical cancer by electrochemotherapy with bleomycin, cisplatin, and calcium: an in vitro experimental study.

The size of the pores created by external electrical pulses is important for molecule delivery into the cell. The size of pores and their distribution on the cell membrane determine the efficiency of molecule transport into the cell. There are very few studies visualizing the presence of electropores. In this study, we aimed to investigate the size distribution of electropores that were created by high intensity and short duration electrical pulses on MCF-7 cell membrane. Scanning Electron Microscopy (SEM) was used to visualize and characterize the membrane pores created by the external electric field. Structural changes on the surface of the electroporated cell membrane was observed by Atomic Force Microscopy (AFM). The size distribution of pore sizes was obtained by measuring the radius of 500 electropores. SEM imaging showed non-uniform patterning. The average radius of the electropores was 12 nm, 51.60% of pores were distributed within the range of 5 to 10 nm, and 81% of pores had radius below 15 nm. These results showed that microsecond (µs) high intensity electrical pulses cause the creation of heterogeneous nanopores on the cell membrane.

The estimation of pore size distribution of electroporated MCF-7 cell membrane.

Colon cancer is particularly increasing in incidence in developed countries, and it ranks first in terms of morbidity and mortality worldwide. As in other types of cancer, metabolic changes and cellular death mechanisms play a critical role in cancer treatment. Cancer cells prefer glycolysis for energy production even in the presence of oxygen; this phenomenon is known as the Warburg effect. Glycolysis, which is essentially an anaerobic process, provides metabolic adaptation by preventing mitochondrial oxidative phosphorylation in cancer cells. Electroporation (EP) is a method that increases the uptake of drugs into the cell by creating temporary pores in the cell membrane. 3-Bromo-2-oxopropionic acid and Lithium chloride (LiCl) are agents that may show anticancer potential through different mechanisms. The aim of this study was to investigate the potential anticancer effects of the EP-assisted combination of 3-Bromo-2-oxopropionic acid and LiCl agents in DLD-1 colon cancer cells and to evaluate how these effects occur at the level of cell viability, apoptotic activity and metabolic markers. In the study, cell viability was evaluated by WST-8 tetrazolium based colorimetric method. Within the scope of analysis of apoptotic activity and metabolic responses; caspase-3, Phosphatidylinositol 3-kinase (PI3K) and Glucose transporter-1 (GLUT-1) levels were determined by ELISA method. While no change in cell viability was observed at 10 and 20 µM doses of 3-Bromo-2-oxopropionic acid, a significant decrease was detected in its combination with EP. IC value was determined as 37.99 ± 2.34 µM in a single application and as 27.33 ± 1.99 µM in its combination with EP. While low doses of LiCl (5 and 10 mM) did not affect cell viability, a significant decrease was detected at doses of 50, 75 and 100 mM. In its combination with EP, a significant loss of viability was observed even at low doses and the IC value decreased from 98.92 ± 0.64 mM to 81.88 ± 1.64 mM. A decrease in IC doses is observed in the treatment with the combination of both 3-Bromo-2-oxopropionic acid and LiCl with EP. The combined treatment enables the same lethal effect on cancer cells with lower drug doses. Caspase-3 activity increased significantly in combined applications with EP. While 3-Bromo-2-oxopropionic acid alone did not change the level of PI3K, it caused a significant increase in its combination with EP. While LiCl caused an increase in the level of PI3K at low doses, a decrease was detected in its combination with EP. Regular partial increases in GLUT-1 level were determined in a dose-dependent manner in 3-Bromo-2-oxopropionic acid treatment alone. A similar increase was determined at 20 and 30 µM doses combined with EP, while a decrease occurred at 40 µM. While no significant increase was detected in GLUT-1 levels compared to the control group in 10 mM LiCl treatment alone and in combination with EP, a significant increase was detected in 50 and 75 mM LiCl doses compared to the control group. In conclusion, 3-Bromo-2-oxopropionic acid and LiCl show anticancer potential by affecting apoptotic and metabolic processes, and their combination with EP increases these effects. In order to explain the effectiveness of these treatment approaches from every perspective, it is recommended to conduct in vitro studies on different colon cancer cell lines and evaluate them with more comprehensive in vivo and clinical studies.

Effects of 3-Bromo-2-oxopropionic acid and LiCl treatment combined with electroporation on apoptotic and metabolic responses in DLD-1 colon cancer cells.

Article	Year
Treatment of cervical cancer by electrochemotherapy with bleomycin, cisplatin, and calcium: an in vitro experimental study. Medical oncology (Northwood, London, England)\| PubMed ID: 36527530	2022
The estimation of pore size distribution of electroporated MCF-7 cell membrane. Electromagnetic biology and medicine\| PubMed ID: 38900674	2024
Effects of 3-Bromo-2-oxopropionic acid and LiCl treatment combined with electroporation on apoptotic and metabolic responses in DLD-1 colon cancer cells. Medical oncology (Northwood, London, England)\| PubMed ID: 40650691	2025

Guney Gursoy

Biography

JoVE Journal Publications

Other Publications