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Cloning of an isoform of integrin-linked kinase (ILK) that is upregulated in HT-144 melanoma cells following TGF-β1 stimulation

Oncogene volume 19, pages 3069–3077 (2000)Cite this article

Abstract

We have shown previously that integrin-linked kinase (ILK) is upregulated in human HT-144 melanoma cells following TGF-β1 stimulation. Using mRNA from TGF-β1 stimulated HT-144 cells and reverse transcriptase polymerase chain reaction, we have isolated a cDNA encoding a protein highly homologous to ILK. Sequencing of the full-length 1359 base pair cDNA and polypeptide translation revealed that this protein, designated ILK-2, differs from the known ILK (hereafter called ILK-1) by only four amino acids, while the cDNA sequence diverges by 102 nucleotides, thus excluding that ILK-2 is an allelic variant of ILK-1. Expression of ILK-2 mRNA was observed in metastatic human HT-144 melanoma and HT-1080 fibrosarcoma cell lines, but not in normal human tissues. Moreover, stimulation of HT-144 cells with TGF-β1, but not with EGF, PDGF-AB or insulin, induced a selective overexpression of ILK-2 mRNA as compared to ILK-1 mRNA. Bacterially-expressed GST/ILK-2 autophosphorylated and labeled myelin basic protein as well as a recombinant GST/β3 integrin cytoplasmic tail peptide. Transfection of either ILK-2 or ILK-1 cDNA into the non-metastatic melanoma cell line SK-Mel-2, expressing exclusively ILK-1, induced anchorage independent cell growth and cell proliferation, as demonstrated by growth in soft agar. Our data provide evidence that ILK-2 is a new isoform of ILK-1 that is expressed in some highly invasive tumor cell lines but not in normal adult human tissues and whose expression is regulated by TGF-β1.

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Acknowledgements

This work was supported by grants from Centre de Recherche Public–Santé (CRP–Santé, Luxembourg), CNRS (France), Fondation Luxembourgeoise contre le Cancer (Luxembourg) and EC Biomed-Project BMH4-CT98-3517. B Janji is a recipient of a fellowship from the Ministère de la Culture, de l'Enseignement Supérieur et de la Recherche, Luxembourg. The data presented were obtained by B Janji as part of his doctoral thesis submitted to the University Pierre et Marie Curie, Paris, France.

The nucleotide sequence reported in this paper has been submitted to the EMBL/GenBank/DDBJ database with the Accession number AJ277481.

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  1. Laboratoire Franco-Luxembourgeois de Recherche Biomédicale (CRP-Santé/CNRS), University Center, L-1511 Luxembourg, Grand-Duchy of Luxembourg

    Bassam Janji, Chantal Melchior, Laurent Vallar & Nelly Kieffer

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Janji, B., Melchior, C., Vallar, L. et al. Cloning of an isoform of integrin-linked kinase (ILK) that is upregulated in HT-144 melanoma cells following TGF-β1 stimulation. Oncogene 19, 3069–3077 (2000). https://doi.org/10.1038/sj.onc.1203640

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