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Human mucin MUC1 RNA undergoes different types of alternative splicing resulting in multiple isoforms

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Abstract

MUC1 is a transmembrane mucin with important functions in normal and transformed cells, carried out by the extracellular domain or the cytoplasmic tail. A characteristic feature of the MUC1 extracellular domain is the variable number of tandem repeats (VNTR) region. Alternative splicing may regulate MUC1 expression and possibly function. We developed an RT-PCR method for efficient isolation of MUC1 mRNA isoforms that allowed us to evaluate the extent of alternative splicing of MUC1 and elucidate some of the rules that govern this process. We cloned and analyzed 21, 24, and 36 isoforms from human tumor cell lines HeLa, MCF7, and Jurkat, respectively, and 16 from normal activated human T cells. Among the 78 MUC1 isoforms we isolated, 76 are new and different cells showed varied MUC1 expression patterns. The VNTR region of exon 2 was recognized as an intron with a fixed 5′ splice site but variable 3′ splice sites. We also report that the 3506 A/G SNP in exon 2 can regulate 3′ splice sites selection in intron 1 and produce different MUC1 short isoform proteins. Furthermore, the SNP A to G mutation was also observed in vivo, during de novo tumor formation in MUC1+/−KrasG12D/+PtenloxP/loxP mice. No specific functions have been associated with previously reported short isoforms. We now report that one new G SNP-associated isoform MUC1/Y-LSP, but not the A SNP-associated isoform MUC1/Y, inhibits tumor growth in immunocompetent but not immunocompromised mice.

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Acknowledgments

This work was supported by National Institutes of Health (CA056103 and CA073743 to O.J.F. and CA163462 to A.M.V), Department of Defense (OC093429 to AMV), National Science Foundation (MCB-0842725 to C.M.), and the Nathan Arenson Fund (O.J.F.).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Immunology, University of Pittsburgh School of Medicine, E1001 Biomedical Science Tower, 200 Lothrop St, Pittsburgh, PA, 15261, USA

    Lixin Zhang, Anda Vlad, Christine Milcarek & Olivera J. Finn

  2. Department of Obstetrics and Gynecology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA

    Anda Vlad

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  1. Lixin Zhang
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  2. Anda Vlad
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Correspondence to Lixin Zhang or Olivera J. Finn.

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Zhang, L., Vlad, A., Milcarek, C. et al. Human mucin MUC1 RNA undergoes different types of alternative splicing resulting in multiple isoforms. Cancer Immunol Immunother 62, 423–435 (2013). https://doi.org/10.1007/s00262-012-1325-2

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