@article {JAIME-MARTINEZ885, author = {LUIS ARTURO JAIME-MARTINEZ and MONICA L. MARTINEZ-PACHECO and LENA RUIZ-AZUARA and CARMEN MEJIA}, title = {BAX But Not BCL2 Is Necessary for Apoptosis in Neuroblastoma Cells Treated With Casiope{\'\i}na{\textregistered} IIIia}, volume = {42}, number = {2}, pages = {885--892}, year = {2022}, doi = {10.21873/anticanres.15546}, publisher = {International Institute of Anticancer Research}, abstract = {Background/Aim: The emerging antineoplastics Casiope{\'\i}nas{\textregistered} induce uncoupling of the respiratory chain, production of reactive oxygen species (ROS), entry of Bax into mitochondria, and exit of Ca2+ and Bcl-2 from them, leading to apoptosis. This study aimed to elucidate whether BAX and BCL2 are necessary for apoptosis. Materials and Methods: We silenced BAX and BCL2 by CRISPR-Cas9, assessed ROS and calcium retention capacity (CRC) by spectrofluorometry, and caspase-3 with inmunoblotting in neuroblastoma (NB) cells and 3T3-L1 fibroblasts treated with cisplatin and Casiope{\'\i}na IIIia (CasIIIia). Results: We observed an increase in O2{\textbullet}{\textendash} production only in BCL2KO NB cells treated with cisplatin (three-fold) and CasIIIia (five-fold), whereas the production of H2O2 in BCL2KO NB cells treated with cisplatin and CasIIIia increased five-fold and three-fold, respectively. The baseline calcium-retention capacity (CRC) was 1.7 relative fluorescence units (RFU) in both cell types. In BAXKO, cisplatin and CasIIIia increased CRC to ~2.3 RFU, and in BCL2KO, they decreased CRC to ~1.1 RFU. We did not detect caspase-3 in BAXKO NB cells. Conclusion: Only BAX is essential for CasIIIia-induced apoptosis.}, issn = {0250-7005}, URL = {https://ar.iiarjournals.org/content/42/2/885}, eprint = {https://ar.iiarjournals.org/content/42/2/885.full.pdf}, journal = {Anticancer Research} }