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(Cluj-Napoca) Photodynamic therapy on experimental tumors and cell cultures.
Romania (FASMR) - University of Medicine and Pharmacy "Iuliu Hatieganu", Cluj-Napoca
Department of Physiology, University of Medicine and Pharmacy “Iuliu Hatieganu”, Cluj-Napoca, Romania
Professor Doctor Simona Clichici
Professor Doctor Simona Clichici, Professor Doctor Filip Gabriela
Type of Research Project
- Basic science
What is the background of the project?
Photodynamic therapy (PDT) is increasingly being recognized as an attractive, alternative treatment modality for cancer. Efficacy is high for small superficial tumors and, except for temporary skin photosensitization, there are no long-term side effects. This is still not a widely accepted alternative treatment for cancer, and ongoing studies still try to find the best combinations possible to make it more efficient for as many types of cancer. Photodynamic therapy (PDT) is a treatment that uses a drug, called a photosensitizer or photosensitizing agent, and a particular type of light. When photosensitizers are exposed to a specific wavelength of light, they produce a form of oxygen that kills nearby cells. In addition, the cytotoxic reactive oxygen species cause damage of microvessels and induce nonspecific immune reaction. In the first step of PDT for cancer treatment, a photosensitizing agent is injected into the bloodstream. The agent is absorbed by cells all over the body but stays in cancer cells longer than it does in normal cells. Approximately 24 to 72 hours after injection, when most of the agent has left normal cells but remains in cancer cells, the tumor is exposed to light. The photosensitizer in the tumor absorbs the light and produces an active form of oxygen that destroys nearby cancer cells. PTD is a noninvasive therapy used most in oncology and it is an adjuvant method with chemotherapy, radiotherapy and surgery.The search for new, third-generation photosensitizers is still ongoing, especially for drugs that can be activated with light of a longer wavelength, which provoke shorter generalized photosensitivity and have better tumor specificity.
What is the aim of the project?
The aim of the project is to quantify the effect of photodynamic therapy mediated by different porphyrins on tumor cells in vitro and in vivo.
What techniques and methods are used?
The in vitro study will be performed on human and murine melanoma cells treated with various porphyrins and the results will be quantified by evaluation of oxidative stress markers (malondialdehyde, glutathione reduced/oxidized ratio, carbonyls protein and antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase). Oxidative stress is defined as the imbalance in the redox characteristics of some cellular environment which can be the result of either biochemical processes leading to the production of reactive species, exposure to damaging agents. The direct quantification of reactive oxygen and nitrogen species (ROS/RNS) will be assessed with mass spectrometry techniques, targeting the cell cultures both collected from serum and directly from tumours. Spectrofluorimetry methods will be used in order to analyse the inflamed tissue. Transcription factors (nuclear factor kappa-light-chain-enhancer of activated B cells or Nf-kB, phosphorylated nuclear factor kappa-light-chain-enhancer of activated B cells or phospho NF-kB, Activator protein 1 or AP-1, signal transducer and activator of transcription or STAT, Hypoxia-inducible factor 1 or HIF-1, melanocyte inducing transcription factor or MITF) will also be analysed through this method. Anti inflammatory enzymes (COX-2, markers of pigmentation and inflammation markers such as interleukin -6 and tumor necrosis factor alpha will be evaluated through the enzyme-linked immunosorbent assay (ELISA). This is a commonly used analytical biochemistry assay that uses a solid-phase enzyme immunoassay (EIA) to detect the presence of a ligand (commonly a protein) in a liquid sample using antibodies directed against the protein to be measured. The tumor necrosis score will be analysed by quantification using staining methods with hematoxylin and eosin. In in vivo study the experiments will be performed on Walker carcinosarcoma transplantated on Wistar rats. The effects will be quantified by determination of oxidative stress markers, matrix metalloproteinases, transcription factors (NF-kB, AP-1) and inflammation (cytokine levels).
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The students will learn how to manipulate the laboratory animals such as laboratory rats, to anesthetize them, to take biopsies from cancerous tissue and to transplant the tumor. Afterwards they will perform the photodynamic therapy on the animals, take the blood samples and extract tumor tissue. In addition, they will also test the effects of photodynamic therapy on tumor by biochemical evaluation of oxidative stress parameters (malondialdehyde, carbonyl proteins, glutathione) using mass spectrometry techniques. They will learn how to use the spectrofluorimeter in order to perform those laboratory tests. The students will be under the supervision of the tutor, and do all of the experiments with their help. The student will also perform the analysis of the data gathered during his placement, together with the supervisor.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Theoretical teaching will be provided by the tutor and supervisors. The students will receive some lectures for a better understanding of the project.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No specific skills required
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Pre-Medical students from the American-British system
- Agostinis; Patrizia & Berg; Kristian & Cengel; Keith & H Foster; Thomas & Girotti; Albert & Gollnick; Sandra & M Hahn; Stephen & Hamblin; Michael & Juzeniene; Asta & Kessel; David & Korbelik; Mladen & Moan; Johan & Mroz; Pawel & Nowis; Dominika & Piette; Jacques & C Wilson; Brian & Golab; Jakub. (2011).PHOTODYNAMIC THERAPY OF CANCER. CA: a cancer journal for clinicians. 61. 250-81. 10.3322/caac.20114.
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