Citotoksični učinak i apoptoza matičnih stanica karcinoma dojke tretiranih novosintetiziranim protutumorskim spojem tieno[2,3-b]piridinom

Objectives: The purpose of this study is to determine the effects of treating breast cancer stem cells with the newly synthesized thieno[2,3-b]pyridine anticancer agent, by focusing on its cytotoxic and apoptotic effects on the investigated human cell line MDA-MB-231. Methods: The MDA-MB-231 triple-negative breast cancer cell line was treated with a newly developed thienopyridine anticancer compound (3-amino-N-(3-chloro-2-methylphenyl)-5- oxo-5,6,7,8-tetrahydrothieno[2,3-b]quinoline-2-carboxamide, 1) to determine its cytotoxic effect on triple negative breast cancer, the type of cell death it causes, and the cancer stem cell percentage after treatment. The 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay was performed, to analyze the cellular metabolic activity and determine the cytotoxic effect. Flow cytometric analysis was used in combination with Annexin-V-FITC and propidium iodide staining to assess the type of cell death after 48h of treatment with compound 1 (2 µM). Furthermore, flow cytometry also provided the percentage of CD44+/CD24- cancer stem cells after treatment. Results: Compound 1 was cytotoxic for breast cancer cells in a dose- and time-dependent manner; cell death occurs mainly by apoptosis. The percent of cancer stem cells decreased four times. Conclusion: Due to its cytotoxic effect on the percentage of triple negative breast cancer stem cells compound 1 can be a potential treatment for triple-negative breast cancer.Ciljevi: Cilj ovog istraživanja je utvrditi učinke tretmana matičnih stanica karcinoma dojke novosintetiziranim protutumorskim spojem tieno [2,3-b] piridinom, s naglaskom na njegove citotoksične i apoptotske učinke na ispitivanu staničnu liniju MDA-MB-231. Materijali i metode: Trostruko negativna stanična linija karcinoma dojke MDA-MB-231 tretirana je novosintetiziranim tieno-piridinskim spojem (3-amino-N-(3-kloro-2-metilfenil)-5-okso-5,6,7,8-tetrahidrotieno[2,3-b]kinolin-2-karboksamid, 1) kako bi se utvrdio citotoksični učinak, tip stanične smrti i postotak matičnih stanica karcinoma nakon tretmana. Napravljen je MTT (3-(4,5-dimetiltiazolil-2)-2,5-difeniltetrazolij bromid) test kako bi se analizirala stanična metabolička aktivnost i utvrdio citotoksični učinak. Protočna citometrija u kombinaciji s bojanjem aneksin-V-FITC-om i propidij jodidom, korištena je kako bi se utvrdio tip stanične smrti nakon 48h tretmana spojem 1 (2 µM). Nadalje, protočnom citometrijom je također utvrđen postotak CD44+/CD24- matičnih stanica karcinoma nakon tretmana. Rezultati: Spoj 1 bio je citotoksičan za stanice karcinoma dojke te citotoksičnost korelira s koncentracijom spoja i vremenom inkubacije. Stanična smrt je nastupila prvenstveno zbog apoptoze. Postotak matičnih stanica karcinoma smanjio se četiri puta. Zaključci: Zbog citotoksičnog učinka na postotak matičnih stanica trostruko negativnog karcinoma dojke, spoj 1 može biti potencijalna terapija za trostruko negativni karcinom dojke