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Crosstalk between Noxa, Bcl-2, and ceramide in mediating p53-dependent apoptosis in Molt-4 human T-cell leukemia

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Abstract

Ionizing radiation induces apoptosis in human Molt-4 leukemia cells in a p53-dependent manner. The tumor suppressor p53 stimulates various downstream targets that presumably trigger, individually or in concert, de novo ceramide synthesis and intrinsic apoptosis via mitochondrial outer membrane permeabilization (MOMP). Among these targets, BH3-only protein Noxa was found to be promptly activated by p53 prior to ceramide accumulation and apoptosis in response to irradiation. To evaluate the relation between Noxa and ceramide in irradiation-induced apoptosis, Noxa was silenced in Molt-4 cells and apoptosis, p53 expression, and ceramide accumulation were assessed in response to irradiation. In the absence of Noxa, irradiation of Molt-4 cells still induced apoptosis in a p53-dependent manner however ceramide levels decreased significantly although they remained higher than untreated control. Upon irradiation, Noxa was found to translocate to the mitochondria where endogenous ceramide accumulation was observed. In contrast, overexpression of Bcl-2, another mitochondrial protein, in Molt-4 cells abolished the endogenous ceramide accumulation and apoptosis. In irradiation-induced, p53-dependent pathways of apoptosis, the pro-apoptotic Noxa represents one of several, yet to be identified, pathways simultaneously triggered by p53 to produce mitochondrial ceramide accumulation and apoptosis. In contrast, Bcl-2 functions as a broader inhibitor of both ceramide accumulation and apoptosis. Altogether, these results indicate that members of the Bcl-2 family differentially regulate ceramide accumulation and reveal the existence of crosstalk between Bcl-2 family members and ceramide in mediating p53-dependent apoptosis in Molt-4 human T-cell leukemia.

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Funding

This research was supported by funding from the Medical Practice Plan (MPP) at AUB-FM and the Lebanese National Center for Scientific Research (LNCSR) to GD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, Faculty of Medicine, American University of Beirut, Beirut, Lebanon

    Hadile Kobeissy, Zeinab Dakdouk, Lina Kozhaya & Ghassan Dbaibo

  2. Department of Biology, Faculty of Sciences, Lebanese University, Hadath, Lebanon

    Rouba Hage-Sleiman

  3. Department of Pediatrics and Adolescent Medicine, Center for Infectious Diseases Research, Faculty of Medicine, American University of Beirut, Beirut, Lebanon

    Ghassan Dbaibo

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  1. Hadile Kobeissy
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  2. Rouba Hage-Sleiman
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  3. Zeinab Dakdouk
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  4. Lina Kozhaya
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  5. Ghassan Dbaibo
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RHS and GD conceived and designed the project. HK, ZD and LK performed the experiments. HK and RHS analyzed the results. HK and RHS drafted the manuscript. RHS and GD revised and finalized the manuscript which was read and approved by all authors.

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Correspondence to Rouba Hage-Sleiman or Ghassan Dbaibo.

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Kobeissy, H., Hage-Sleiman, R., Dakdouk, Z. et al. Crosstalk between Noxa, Bcl-2, and ceramide in mediating p53-dependent apoptosis in Molt-4 human T-cell leukemia. Mol Cell Biochem 475, 215–226 (2020). https://doi.org/10.1007/s11010-020-03874-9

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