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Caspase cleavage of Mcl-1 impairs its anti-apoptotic activity and proteasomal degradation in non-small lung cancer cells

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Abstract

Global cleavage of cellular proteins by activated caspases is a hallmark of apoptosis, which causes biochemical collapse of the cell. Recent studies suggest that, rather than completely destroying a protein, caspase cleavage can confer novel characteristics or functions. In this respect, the post-caspase role of Bcl-2 family proteins remains uncharacterized. Here, we showed that Mcl-1, a pro-survival member of the Bcl-2 family, was cleaved by caspase-3 in non-small cell lung cancer (NSCLC) cells undergoing chemotherapeutic agent-triggered apoptosis. Caspase cleavage partially impaired the anti-apoptotic activity of Mcl-1 by reducing its mitochondrial localization and impeding its association with the permeability transition pore-forming protein Bak. However, the stability of cleaved Mcl-1 was markedly enhanced because it was more refractory to ubiquitination-dependent proteasomal degradation, thereby improving cell viability to a greater extent than full-length Mcl-1 when transiently expressed in NSCLC cells. These findings shed new light on the role of Mcl-1 in apoptosis and suggest potential novel targets for optimizing the tumoricidal capacity of chemotherapy.

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Abbreviations

Mcl-1:

Myeloid cell leukemia 1

BH:

Bcl-2 homology

Bax:

Bcl-2-associated protein X

NSCLC:

Non-small cell lung cancer

PT:

Permeability transition

PARP:

Poly ADP-ribose polymerase

CHX:

Cyclohexane

CCK-8:

Cell Counting Kit-8

PI:

Propidium iodide

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Nos. 81472631 and 31200429).

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Author notes

  1. Ting Wang, Zhiwei Yang and Yimeng Zhang have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, **’an, 710032, China

    Ting Wang, Yimeng Zhang, **ang Zhang, Lei Wang & Lintao Jia

  2. Department of Applied Physics, **’an Jiaotong University, **’an, 710049, China

    Zhiwei Yang & Shengli Zhang

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  1. Ting Wang
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Correspondence to Shengli Zhang or Lintao Jia.

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Electronic supplementary material

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10495_2017_1436_MOESM1_ESM.tif

Supplementary Figure 1. Survival of lung cancer patients (Total) and those receiving routine chemotherapy as grouped by Mcl-1 expression levels. (TIF 98 KB)

10495_2017_1436_MOESM2_ESM.tif

Supplementary Figure 2. Western blot analysis of cells after treatment with CP (20 μmol/L) for the indicated times. (TIF 57 KB)

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Wang, T., Yang, Z., Zhang, Y. et al. Caspase cleavage of Mcl-1 impairs its anti-apoptotic activity and proteasomal degradation in non-small lung cancer cells. Apoptosis 23, 54–64 (2018). https://doi.org/10.1007/s10495-017-1436-5

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