Intrinsic and extrinsic apoptosis responses in leukaemia cells following daunorubicin treatment

Abstract

Background Daunorubicin is used clinically in the treatment of myeloma, acute lymphatic and myelocytic leukaemia. The toxic lesions caused by daunorubicin induce various modes of cell death, including apoptosis. Apoptosis is highly regulated programmed cell death that can be initiated mainly via two pathways, through death receptors (extrinsic) or involvement of the mitochondria (intrinsic). Induction of apoptosis via these pathways has been alluded following treatment with daunorubicin, but never compared in acute lymphoblastic leukaemia over a time course. Methods This study investigated the mechanisms of daunorubicin induced apoptosis in the treatment of CCRF-CEM, MOLT-4 (acute T-lymphoblastic leukaemia) and SUP-B15 (acute B-lymphoblastic leukaemia) cells. Cells were treated with daunorubicin for 4 h, and then placed in recovery medium (without daunorubicin) for 4 h, 12 h and 24 h. Apoptotic response was analysing using annexin-V expression, caspase activity, mitochondrial membrane potential change and an array to detect 43 apoptotic proteins. Results Daunorubicin induced apoptosis in all leukemic cell lines, but with different levels and duration of response. Both apoptosis levels and caspase activity increased after four hours recovery then declined in CCRF-CEM and MOLT-4 cells. However, SUP-B15 cells displayed initially comparable levels but remained elevated over the 24 h assessment period. Changes in mitochondrial membrane potential occurred in both MOLT-4 and CCRF-CEM cells but not in SUP-B15 cells. Expression of apoptotic proteins, including Bcl-2, Bax, caspase 3 and FADD, indicated that daunorubicin potentially induced both extrinsic and intrinsic apoptosis in both CCRF-CEM and MOLT-4 cells, but only extrinsic apoptosis in SUP-B15 cells. Conclusions This study describes variations in sensitivities and timing of apoptotic responses in different leukaemia cell lines. These differences could be attributed to the lack of functional p53 in coordinating the cells response following cytotoxic treatment with daunorubicin, which appears to delay apoptosis and utilises alternative signalling mechanisms that need to be further explored.

Platelet Microparticles Decrease Daunorubicin-Induced DNA Damage and Modulate Intrinsic Apoptosis in THP-1 Cells

“…The cytotoxic effect of DNR is commonly associated with an increase in DNA damage, i.e., an intrinsic stimulus. However, it is also suggested to activate the extrinsic apoptotic pathway [ 55 ]. To evaluate activation of intrinsic and extrinsic apoptosis after DNR-treatment, we measured levels of active caspase-8 and caspase-9 by flow cytometry, and gated the cells in “lo”, “mid”, and “hi” populations.…”

“…The dependence of intact p53 protein for apoptosis induction by doxorubicin, a related Top2 poison, suggests a strong reliance on the activation of the intrinsic apoptotic pathway for this class of chemotherapeutics [ 66 ]. However, there is evidence that DNR-treatment upregulates death receptors and activate caspase-8 in multiple leukemic cell lines, thereby inducing extrinsic apoptosis [ 55 ]. In our experiments, caspase-8 and caspase-9 activation was equally inhibited by PMPs, suggesting that PMPs interfere with a common activation mechanism.…”

Knockdown of microRNA-375 suppresses cell proliferation and promotes apoptosis in human breast cancer cells

“…Generally, cells commit to die due to their own pro-anti apoptotic proteins in apoptosis there by, it is generally referred to as a cell suicidal process and the balance between oncoproteins and tumour suppressor proteins are more responsible to determine when the cells die. So, this apoptotic mechanism is accomplished by both intrinsic (mitochondrial) and extrinsic (death receptor) pathway signaling protein molecules activation (49,50) .…”

The anticarcinogenic and anticancer effects of the dietary flavonoid, morin: Current status, challenges, and future perspectives

“…The major role of anti-apoptotic proteins is to directly interact with pro-apoptotic proteins and suppress their activity, leading to the inhibition of apoptosis (Czabotar, Lessene, Strasser, & Adams, 2014;Kale, Osterlund, & Andrews, 2018). The extrinsic apo- cell death (Al-Aamri et al, 2021;McIlwain, Berger, & Mak, 2013). During apoptosis process, activated initiator caspases (caspase-2, caspase-8, caspase-9, and caspase-10) cleave and activate the downstream executioner caspases (caspase-3, caspase-6, and caspase-7) that cause apoptosis by degrading cellular components (Parrish, Freel, & Kornbluth, 2013;Patel, Balakrishnan, Keating, Wierda, & Gandhi, 2015;Vigneswara & Ahmed, 2020).…”

“…The major role of anti‐apoptotic proteins is to directly interact with pro‐apoptotic proteins and suppress their activity, leading to the inhibition of apoptosis (Czabotar, Lessene, Strasser, & Adams, 2014; Kale, Osterlund, & Andrews, 2018). The extrinsic apoptosis pathway is activated by interacting death ligands such as Fas ligand (FasL), Tumor necrosis factor ( TNF ), TNF‐related apoptosis ‐inducing ligand (TRAIL) with death receptors, including Fas (also known as CD95), TRAIL receptor (TRAIL‐R) or tumor necrosis factor receptor 1 (TNFR1) (Al‐Aamri, Irving, Bradley, & Meehan‐Andrews, 2021; Green & Llambi, 2015). Both pathways activate the caspase protease family, which acts in a proteolytic cascade to induce cell death (Al‐Aamri et al, 2021; McIlwain, Berger, & Mak, 2013).…”