Recent studies demonstrated that 2MM-67, a new 3-(5-imidazo[2,1-b]thiazolylmethylene)-2-indolinone, analog of recently published compounds is able to induce apoptotic cell death in HT-29, a colon carcinoma cell line. The cells treated with the proapoptotic agent (500 nM) showed arrest in G2/M phase and increased expression of p21 protein. Moreover it was observed a significant reduction of mitochondrial potential (Δψm), enhanced expression of Bax and, finally, a marked induction of effector caspases. The aim of this work was to assess cellular magnesium levels during 2MM-67-induced apoptosis and if they correlate to Δψm decrease and to an increase in ROS production, characteristic feature of mitochondrion-mediated apoptosis. Intracellular free and total magnesium were evaluated in flow cytometry, respectively by the commercial probe MagFluo4-AM (Kd=4.7 mM), and the new synthesized DCHQ7 (Kd=8.3 μM), while ROS production was assessed by DiClFDA and Dihydrohetidine. Our results indicate that 2MM-67 treatment in HT29 cells can result in rapid declines in intracellular free and total magnesium concomitant with the decrease in mitochondrial membrane potential and the increase of ROS production.
A new imidazothiazolylmethylen-oxindole induces apoptosis and decline in intracellular free and total magnesium in HT29 colon cancer cells
CAPPADONE, CONCETTINA;FARRUGGIA, GIOVANNA;MARRACCINI, CHIARA;SGARZI, MASSIMO;LOMBARDO, MARCO;IOTTI, STEFANO
2011
Abstract
Recent studies demonstrated that 2MM-67, a new 3-(5-imidazo[2,1-b]thiazolylmethylene)-2-indolinone, analog of recently published compounds is able to induce apoptotic cell death in HT-29, a colon carcinoma cell line. The cells treated with the proapoptotic agent (500 nM) showed arrest in G2/M phase and increased expression of p21 protein. Moreover it was observed a significant reduction of mitochondrial potential (Δψm), enhanced expression of Bax and, finally, a marked induction of effector caspases. The aim of this work was to assess cellular magnesium levels during 2MM-67-induced apoptosis and if they correlate to Δψm decrease and to an increase in ROS production, characteristic feature of mitochondrion-mediated apoptosis. Intracellular free and total magnesium were evaluated in flow cytometry, respectively by the commercial probe MagFluo4-AM (Kd=4.7 mM), and the new synthesized DCHQ7 (Kd=8.3 μM), while ROS production was assessed by DiClFDA and Dihydrohetidine. Our results indicate that 2MM-67 treatment in HT29 cells can result in rapid declines in intracellular free and total magnesium concomitant with the decrease in mitochondrial membrane potential and the increase of ROS production.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
