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CACNA2D2-mediated apoptosis in NSCLC cells is associated with alterations of the intracellular calcium signaling and disruption of mitochondria membrane integrity

Oncogene volume 22, pages 615–626 (2003)Cite this article

Abstract

The CACNA2D2 gene, a new subunit of the Ca2+-channel complex, was identified in the homozygous deletion region of chromosome 3p21.3 in human lung and breast cancers. Expression deficiency of the CACNA2D2 in cancer cells suggests a possible link of it to Ca2+ signaling in the pathogenesis of lung cancer and other cancers. We investigated the effects of overexpression of CACNA2D2 on intracellular Ca2+ contents, mitochondria homeostasis, cell proliferation, and apoptosis by adenoviral vector-mediated wild-type CACNA2D2 gene transfer in 3p21.3-deficient nonsmall cell lung cancer cell lines. Exogenous expression of CACNA2D2 significantly inhibited tumor cell growth compared with the controls. Overexpression of CACNA2D2 induced apoptosis in H1299 (12.5%), H358 (13.7%), H460 (22.3%), and A549 (50.1%) cell lines. Levels of intracellular free Ca2+ were elevated in AdCACNA2D2-transduced cells compared with the controls. Mitochondria membrane depolarization was observed prior to apoptosis in Ad-CACNA2D2 and Adp53-transduced H460 and A549 cells. Release of cyt c into the cytosol, caspase 3 activation, and PARP cleavage were also detected in these cells. Together, these results suggest that one of the pathways in CACNA2D2-induced apoptosis is mediated through disruption of mitochondria membrane integrity, the release of cyt c, and the activation of caspases, a process that is associated with regulation of cytosolic free Ca2+ contents.

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Abbreviations

ADP:

adenosinediphosphate

CACNA2D2:

calcium-channel alfa-2-delta-2 subunit

COX IV (I):

cytochrome oxidase IV subunit I

cyt C:

cytochrome C

DAPK:

death-associated protein kinase

DMSO:

dimethylsulfoxide

FBS:

fetal bovine serum

HBSS:

Hanks balanced saline solution

MOI:

multiplicity of infection

NSCLC:

non-small cell lung cancer

PARP:

poly ADP-ribose polymerase

PI:

propidiumiodide

TUNEL:

terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling

VACC:

voltage-activated calcium channel

XTT:

sodium 3,3′-{1-[(phenylamino)carbonyl]-3,4-tetrazolium}-bis(4-methoxy-6-nitro)-benzene sulfonic acid hydrate

wt:

wild-type

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Acknowledgements

The authors would like to thank Karen Ramirez and Wendy Schober-Ditmore for their assistance in FACS analysis, and David McConkey, Leta Nutt, and Abujiang Pataer for discussions on the methodology. This work was partially supported by grants from the National Cancer Institute, the National Institutes of Health SPORE (2P50-CA70907-04); (P01 CA78778-01A1) (JAR); (CA71618) (JDM), a WM Keck Gene Therapy Career Development Grant (LJ), by a grant from the Department of the Army BESCT Lung Cancer Program (DAMD17011068902); by the Swiss National Science Foundation (GLC) and Bernische Krebsliga (GLC); by gifts to the Division of Surgery MD Anderson Cancer Center, from Tenneco and Exxon for the Core Laboratory Facility; by the M. D. Anderson Cancer Center Support Core Grant (CA16672); by a grant from the Tobacco Settlement Funds as appropriated by the Texas State Legislature (Project 8), and by a sponsored research agreement with Introgen Therapeutics, Inc. (SR93-004-1).

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Authors and Affiliations

  1. Department of Thoracic and Cardiovascular Surgery, Section of Thoracic Molecular Oncology, The University of Texas MD Anderson Cancer Center, Houston, 77030, TX, USA

    Giovanni L Carboni, Masahiko Nishizaki, Kai Xu, Jack A Roth & Lin Ji

  2. Department of Internal Medicine and Pharmacology, Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, 75390, TX, USA

    Boning Gao & John D Minna

  3. Division of General Thoracic Surgery, University Hospital, Bern, Switzerland

    Giovanni L Carboni

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  1. Giovanni L Carboni
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Correspondence to Lin Ji.

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Carboni, G., Gao, B., Nishizaki, M. et al. CACNA2D2-mediated apoptosis in NSCLC cells is associated with alterations of the intracellular calcium signaling and disruption of mitochondria membrane integrity. Oncogene 22, 615–626 (2003). https://doi.org/10.1038/sj.onc.1206134

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