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Selective regulation of p38β protein and signaling by integrin-linked kinase mediates bladder cancer cell migration

Oncogene volume 33, pages 690–701 (2014)Cite this article

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Abstract

Integrin-linked kinase (ILK) and p38MAPK are protein kinases that transduce extracellular signals regulating cell migration and actin cytoskeletal organization. ILK-dependent regulation of p38MAPK is critical for mammalian kidney development and in smooth muscle cell migration, however, specific p38 isoforms has not been previously examined in ILK-regulated responses. Signaling by ILK and p38MAPK is often dysregulated in bladder cancer, and here we report a strong positive correlation between protein levels of ILK and p38β, which is the predominant isoform found in bladder cancer cells, as well as in patient-matched normal bladder and tumor samples. Knockdown by RNA interference of either p38β or ILK disrupts serum-induced, Rac1-dependent migration and actin cytoskeletal organization in bladder cancer cells. Surprisingly, ILK knockdown causes the selective reduction in p38β cellular protein level, without inhibiting p38β messenger RNA (mRNA) expression. The loss of p38β protein in ILK-depleted cells is partially rescued by the 26S proteasomal inhibitor MG132. Using co-precipitation and bimolecular fluorescent complementation assays, we find that ILK selectively forms cytoplasmic complexes with p38β. In situ proximity ligation assays further demonstrate that serum-stimulated assembly of endogenous ILK–p38β complexes is sensitive to QLT-0267, a small molecule ILK kinase inhibitor. Finally, inhibition of ILK reduces the amplitude and period of serum-induced activation of heat shock protein 27 (Hsp27), a target of p38β implicated in actin cytoskeletal reorganization. Our work identifies Hsp27 as a novel target of ILK–p38β signaling complexes, playing a key role in bladder cancer cell migration.

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Acknowledgements

We thank Drs Aaron Irving, Dakang Xu and Howard Yim (Centre for Cancer Research, MIMR) for consultation and helpful discussions. Dr S Dedhar (University of British Columbia) kindly provided QLT-0267. This work was supported by grants to GH and EDW from the National Health and Medical Research Council of Australia, with additional support through the Victorian Government’s Operational Infrastructure Support Program.

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  1. L Yu and X Yuan: These authors contributed equally to this work.

  2. E D Williams: Co-senior corresponding author.

Authors and Affiliations

  1. Centre for Cancer Research, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia

    L Yu, D Wang, B Barakat, E D Williams & G E Hannigan

  2. Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiaotong, University School of Medicine, Shanghai, China

    X Yuan

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Correspondence to E D Williams or G E Hannigan.

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Yu, L., Yuan, X., Wang, D. et al. Selective regulation of p38β protein and signaling by integrin-linked kinase mediates bladder cancer cell migration. Oncogene 33, 690–701 (2014). https://doi.org/10.1038/onc.2013.20

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