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MMP-2 siRNA induced Fas/CD95-mediated extrinsic II apoptotic pathway in the A549 lung adenocarcinoma cell line

Oncogene volume 26, pages 7675–7683 (2007)Cite this article

Abstract

We have previously reported that the downregulation of MMP-2 by adenovirus-mediated delivery of MMP-2 siRNA (Ad-MMP-2) reduced spheroid invasion and angiogenesis in vitro, and, metastasis and tumor growth in vivo. In this study, we investigated the mechanism of Ad-MMP-2-mediated growth inhibition in vitro and in vivo. Ad-MMP-2 infection led to the induction of apoptosis as determined by TUNEL assay, Annexin-V staining and PARP-1 cleavage in a dose-dependent manner in A549 cells. Ad-MMP-2 decreased the content of the antiapoptotic members of the Bcl-2 family proteins (Bcl-2 and Bcl-xL) and increased the content of the pro-apoptotic members of the Bcl-2 family (Bax and Bcl-xS) as determined by immunoblotting analysis. Furthermore, Ad-MMP-2-mediated apoptosis was accompanied by increase in truncated Bid, release of cytochrome c and the activation of caspase-8, -9 and -3. Immunoblot analysis showed that Ad-MMP-2 infection caused upregulation of Fas/Fas-L and FADD, and Anti-Fas-L antibody reversed Ad-MMP-2-induced apoptosis. Tissue inhibitor of metalloproteinases (TIMP)-3, an endogenous inhibitor of MMP-2, which cleaves Fas-L and activates the Fas/Fas-L inducing apoptotic pathway, was increased in Ad-MMP-2-treated cells. Adenovirus-mediated expression of MMP-2 siRNA in human lung xenografts in vivo resulted in increased immunostaining of Fas, Fas-L, cleaved Bid and TIMP-3. This is the first report, to our knowledge, showing that MMP-2 inhibition upregulates TIMP-3 levels, which in turn, promotes apoptosis in lung cancer.

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Acknowledgements

We thank Noorjehan Ali for technical assistance, Shellee Abraham for manuscript preparation and Diana Meister and Sushma Jasti for review of this paper. This research was supported by National Cancer Institute Grant CA75557, CA92393, CA95058, CA116708 and NINDS NS47699, NS57529 and Caterpillar Inc., OSF Saint Francis Medical Center, Peoria, IL, USA (to JSR) and American Cancer Society Grant no.; 06-03 (SSL).

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  1. Department of Cancer Biology and Pharmacology, Program of Cancer Biology, University of Illinois College of Medicine at Peoria, Peoria, IL, USA

    C Chetty, P Bhoopathi, S S Lakka & J S Rao

  2. Department of Neurosurgery, University of Illinois College of Medicine at Peoria, Peoria, IL, USA

    J S Rao

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  1. C Chetty
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  2. P Bhoopathi
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Correspondence to J S Rao.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Chetty, C., Bhoopathi, P., Lakka, S. et al. MMP-2 siRNA induced Fas/CD95-mediated extrinsic II apoptotic pathway in the A549 lung adenocarcinoma cell line. Oncogene 26, 7675–7683 (2007). https://doi.org/10.1038/sj.onc.1210584

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