Učinak kvercetina na vijabilnost MDA-MB-231 stanica karcinoma dojke

International audienceBreast cancer is the most common malignant disease among women. Approximately 2500 women are diagnosed annually in the Republic of Croatia and more than 900 of them die. Breast cancer therapy depends on the stage and the molecular type of the disease. Major challenge is a triple negative breast cancer.Except for conventional options including surgical treatment, radiotherapy or chemotherapy,there is no specific treatment target. MDA-MB-231 is a breast cancer cell line with negative expression of estrogen, progesterone and HER2 (human epidermal growth factor 2) receptors. It is characterized by its aggressive nature,high degree of proliferation and rapid growth with a high tendency of metastasis resulting in a poor prognosis. According to the latest findings, interest in the context of prevention and antitumour therapy is directed to natural flavonoid compounds. Bioflavonoids are polyphenolic compounds with positive effects on human health. Quercetin is bioflavonoid with a wide spectrum of biological and pharmacological activities. In addition to significant anti-inflammatory and antioxidant activity, quercetin has anticarcinogenic and antiproliferative potential. It is proven to slow down tumour growth and progression of various types of cancer. Tumor cells reprogram energy metabolism and consume large amounts of glucose by overexpressing GLUT transporter for glucose uptake into the cell. Quercetin acts as an inhibitor of GLUT1 transporter, thus reducing the ability of malignant cells to utilize glucose by glycolysis which affects invasiveness. Quercetin also has proapoptotic effect on malignant breast cancer cells. The aim of this experiment was to investigate the effect of quercetin on the viability or metabolic activity, morphology and apoptosis of MDA-MB-231 breast cancer cells at different bioenergy conditions defined by glucose concentrations (0, 1, 5, 10, 25 and 50 mM). MTT assay was performed and the results showed that at all glucose concentrations, quercetin caused a decrease in tumor cell viability after 24-hour and 48-hour exposure.After staining MDA-MB-231 cell preparations with hematoxylin and eosin and microscopy, quercetin has been shown to affect number and morphological characteristics of cells. The Apoptosis detection was performed by TUNEL method (Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) with fluorescence microscopy analysis. However, in these bioenergy conditions and the tested concentration of 100 mM quercetin, apoptosis was not detected. Based on the preliminary results, the cytotoxic and anti-tumor activity of the bioflavonoid quercetin on the MDA-MB-231 breast cancer cells is observed and this experiment can serve as a basis for further experiments of quercetin mechanisms on tumor cells.Karcinom dojke najčešća je maligna bolest žena današnjice od koje u Republici Hrvatskoj godišnje obolijeva oko 2500 žena, a umire njih više od 900. Terapija karcinoma dojke provodi se ovisno o stadiju i molekularnoj vrsti bolesti. Veliki izazov predstavlja trostruko negativan karcinom dojke za koji ne postoji specifična meta liječenja izuzev konvencionalnih opcija liječenja kirurškim zahvatom, radioterapijom ili kemoterapijom. MDA-MB-231 stanična je linija karcinoma dojke koji je negativan na ekspresiju receptora estrogenskog, progesteronskog i HER2 humanog epidermalnog faktora rasta. Karakterizira ga agresivnost, visok stupanj proliferacije i brzog rasta sa tendencijom metastaziranja što rezultira i lošijom prognozom. Prema najnovijim spoznajama, interes u kontekstu prevencije i antitumorske terapije usmjerava se na prirodne spojeve flavonoide. Bioflavonoidi su polifenolni spojevi sa pozitivnim učincima na zdravlje čovjeka. Kvercetin je predstavnik bioflavonoida sa širokim spektrom biološke i farmakološke aktivnosti.Pored značajne antiupalne i antioksidativne aktivnosti, kvercetin djeluje antikancerogeno i antiproliferativno usporavajući razvoj tumora i napredak različitih vrsta karcinoma. Tumorske stanice reprogramiraju svoj energetski metabolizam i troše velike količine glukoze pojačano eksprimirajući GLUT transporter za unos glukoze u stanicu. Kvercetin djeluje kao inhibitor GLUT1 transportera reducirajući tako mogućnost malignih stanica za iskorištenje glukoze glikolizom. Kvercetin ima i proapoptotičko djelovanje na maligne stanice karcinoma dojke. Cilj ovog rada bio je ispitati učinak kvercetina na vijabilnost, odnosno metaboličku aktivnost, morfologiju i apoptozu MDA-MB-231 stanica karcinoma dojke pri različitim bioenergetskim uvjetima, odnosno koncentracijama glukoze (0, 1, 5, 10, 25 i 50 mM). Provođenjem MTT testa, pri svim koncentracijama glukoze pokazalo se da kvercetin uzrokuje pad vijabilnosti tumorskih stanica nakon 24-satne i 48-satne izloženosti. Nakon bojanja MDA-MB-231 staničnih preparata hematoksilinom i eozinom i mikroskopiranja pokazalo se da kvercetin utječe na broj i morfološke karakteristike stanica. Provođena je detekcija apoptoze TUNEL metodom (engl. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) uz analizu fluorescentnom mikroskopijom. Međutim, u navedenim bioenergetskim uvjetima i ispitanoj koncentraciji kvercetina od 100 mM nije zabilježena. Na temelju rezultata može se prepostaviti citotoksično i antitumorsko djelovanje bioflavonoida kvercetina na MDA-MB-231 stanice karcinoma dojke i može poslužiti kao osnova za daljnja ispitivanja mehanizama kvercetina na tumorske stanice

Développement d'aptamères pour la détection de biomarqueurs de cancers ovariens dans les urines

Ovarian cancer is the deadliest gynecological cancer in the world. Early detection is the key for most cancers, but for ovarian cancer, late diagnosis remains one of the main obstacles to recovery and survival of patients. Currently, there is no specific and sufficiently sensitive diagnostic test for the disease at an early stage. The objective of the CARIBOU project (CAncer ovaRIan BiosensOr in Urine) is the development of high-affinity nucleic acid aptamers for the detection of cancer biomarkers in urine. The new method used to obtain the detection part of the biosensor, High-Fidelity Systematic Evolution of Ligands by EXponential enrichment (Hi-Fi SELEX), is used for the selection of aptamers that specifically and sensitively target biomarkers. Cancer biomarkers present in urine are stable, allowing easy and homogenous detection. Ultimately, the use of aptamers as diagnostic tools in biosensor would make it possible to develop a non-invasive test for systematic first-line use.Le cancer de l'ovaire est le cancer gynécologique le plus mortel dans le monde. La détection précoce est la clé pour la plupart des cancers, mais pour le cancer de l'ovaire, le diagnostic tardif reste l'un des principaux obstacles au rétablissement et à la survie des patientes avec un taux de mortalité très élevé. À l'heure actuelle, il n'existe pas de test de diagnostic spécifique et suffisamment sensible pour la maladie à un stade précoce. L'objectif du projet CARIBOU (CAncer ovaRIan BiosensOr in Urine) est le développement d'aptamères d'acides nucléiques de haute spécificité pour la détection de biomarqueurs du cancer dans l'urine. La nouvelle méthode utilisée pour obtenir la partie détection du biocapteur, High-Fidelity Systematic Evolution of Ligands by EXponential enrichment (Hi-Fi SELEX), est utilisée pour la sélection d'aptamères qui ciblent spécifiquement et avec sensibilité les biomarqueurs. Les biomarqueurs du cancer présents dans l'urine sont stables, permettant une détection facile et homogène. A terme, l'utilisation des aptamères comme outils de diagnostic en biocapteur permettrait de développer un test non-invasif pour une utilisation systématique en première ligne

Development of the aptamers for detection of ovarian cancer biomarkers in urine

Le cancer de l'ovaire est le cancer gynécologique le plus mortel dans le monde. La détection précoce est la clé pour la plupart des cancers, mais pour le cancer de l'ovaire, le diagnostic tardif reste l'un des principaux obstacles au rétablissement et à la survie des patientes avec un taux de mortalité très élevé. À l'heure actuelle, il n'existe pas de test de diagnostic spécifique et suffisamment sensible pour la maladie à un stade précoce. L'objectif du projet CARIBOU (CAncer ovaRIan BiosensOr in Urine) est le développement d'aptamères d'acides nucléiques de haute spécificité pour la détection de biomarqueurs du cancer dans l'urine. La nouvelle méthode utilisée pour obtenir la partie détection du biocapteur, High-Fidelity Systematic Evolution of Ligands by EXponential enrichment (Hi-Fi SELEX), est utilisée pour la sélection d'aptamères qui ciblent spécifiquement et avec sensibilité les biomarqueurs. Les biomarqueurs du cancer présents dans l'urine sont stables, permettant une détection facile et homogène. A terme, l'utilisation des aptamères comme outils de diagnostic en biocapteur permettrait de développer un test non-invasif pour une utilisation systématique en première ligne.Ovarian cancer is the deadliest gynecological cancer in the world. Early detection is the key for most cancers, but for ovarian cancer, late diagnosis remains one of the main obstacles to recovery and survival of patients. Currently, there is no specific and sufficiently sensitive diagnostic test for the disease at an early stage. The objective of the CARIBOU project (CAncer ovaRIan BiosensOr in Urine) is the development of high-affinity nucleic acid aptamers for the detection of cancer biomarkers in urine. The new method used to obtain the detection part of the biosensor, High-Fidelity Systematic Evolution of Ligands by EXponential enrichment (Hi-Fi SELEX), is used for the selection of aptamers that specifically and sensitively target biomarkers. Cancer biomarkers present in urine are stable, allowing easy and homogenous detection. Ultimately, the use of aptamers as diagnostic tools in biosensor would make it possible to develop a non-invasive test for systematic first-line use

Detection of the ovarian cancer biomarker in body fluids using nucleic acid aptamers as diagnostic probes

EACR 2022 - Innovative Cancer Science: Translating Biology to MedicineInternational audienc

Detection of the ovarian cancer biomarker in body fluids using nucleic acid aptamers as diagnostic probes

EACR 2022 - Innovative Cancer Science: Translating Biology to MedicineInternational audienc