Indukcija programirane stanične smrti djelovanjem odabranih izotiocijanata

Cilj istraživanja je bio dokazati citotoksični i apoptotički učinak izotiocijanata (ITC): 2-metoksifenila, 4-(metilsulfanil)fenila i 4-nitrofenila na staničnim linijama trostruko-negativnog karcinoma dojke MDA-MB-231 i karcinoma mokraćnog mjehura T24. Pretpostavka je da će se nakon tretiranja stanica spojevima smanjiti postotak metaboličkih aktivnih stanica te da navedeni spojevi uzrokuju programiranu staničnu smrt ispitanih stanica. Ispitivanje citotoksičnosti na staničnim linijama karcinoma dojke MDA-MB-231 i karcinoma mokraćnog mjehura T24 provedeno je MTT testom. Metabolička aktivnost stanica se određivala unutar 4, 24, 48 i 72 h inkubacije. Rezultati za test citotoksičnosti su prikazani grafovima na kojima su prikazan postotak metabolički aktivnih stanica u ovisnosti o koncentraciji spoja i vremenu izlaganja. Svi spojevi pokazuju najveću citotoksičnu aktivnost u koncentracijama 50 µg/mL i 100 µg/mL i u vremenu inkubacije od 72 h, a najbolju citotoksičnost pokazuje spoj 4-(metilsulfanil)fenil izotiocijanat. Bolji citotoksični učinak izotiocijanata je dokazan na staničnoj liniji T24. Nakon toga je ispitan učinak na apoptozu metodom protočne citometrije korištenjem Annexin-V/PI kita. Tretman stanica koncentracijama jednakim IC50 vrijednostima za 48 h inkubacije dokazao je da je mehanizam citotoksične aktivnosti za sva tri uzorka izotiocijanata bio poticanje apoptoze. 2-metoksifenil ITC pokazuje najbolji učinak na poticanje apoptoze na staničnoj liniji MDA-MB-231. 4-nitrofenil ITC pokazuje najbolji učinak na poticanje apoptoze na staničnoj liniji T24.
The aim of the study was to demonstrate the cytotoxic and apoptotic effect of isothiocyanates (ITC): 2-methoxyphenyl, 4- (methylsulfanyl) phenyl and 4-nitrophenyl on MDA-MB-231 triple-negative breast cancer cell lines and T24 bladder cancer cell line. It is assumed that after treatment of cells with compounds, the percentage of metabolically active cells will decrease and that these compounds cause programmed cell death of the tested cells. Cytotoxicity testing on MDA-MB-231 breast cancer cell lines and T24 bladder cancer was performed by MTT assay. Metabolic activity of the cells was determined within 4, 24, 48 and 72 h of incubation. The results for the cytotoxicity test are shown by graphs showing metabolically active cells as a function of compound concentration and exposure time. All compounds showed the highest cytotoxic activity at concentrations of 50 µg/mL and 100 µg/mL and at an incubation time of 72 h, and the best cytotoxicity was shown by the compound 4-(methylsulfanyl) phenyl ITC. A better cytotoxic effect of ITC was demonstrated on the T24 cell line. The effect on apoptosis was then examined by flow cytometry using the Annexin-V / PI kit. Treatment of cells with concentrations equal to IC50 values for 48 h of incubation proved that the mechanism of cytotoxic activity for all three isothiocyanate samples was the induction of apoptosis. 2-methoxyphenyl ITC shows the best effect on inducing apoptosis on the MDA-MB-231 cell line. 4-nitrophenyl ITC shows the best effect on inducing apoptosis on the T24 cell line.