Identification of alternatively spliced TIMP-1 mRNA in cancer cell lines and colon cancer tissue

Usher, P.A.; Sieuwerts, A.M.; Bartels, A.; Lademann, U.; Nielsen, H.J.; Holten-Andersen, L.; Foekens, J.A.; Brünner, N.; Offenberg, H.

doi:10.1016/j.molonc.2007.05.002

« Previous Next »Molecular Oncology
Volume 1, Issue 2 , Pages 205-215, September 2007

Identification of alternatively spliced TIMP-1 mRNA in cancer cell lines and colon cancer tissue

P.A. Usher
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
A.M. Sieuwerts
- Department of Medical Oncology, Erasmus Medical Centre, Rotterdam, The Netherlands
A. Bartels
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
U. Lademann
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
H.J. Nielsen
- Department of Surgical Gastroenterology, University Hospital Hvidovre, Denmark
L. Holten-Andersen
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
J.A. Foekens
- Department of Medical Oncology, Erasmus Medical Centre, Rotterdam, The Netherlands
N. Brünner
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
H. Offenberg
- Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Ridebanevej 9, 1870 Frederiksberg C, Denmark
- Corresponding author. Tel.: +45 35283171; fax: +45 35353514.

Received 15 January 2007; received in revised form 2 May 2007; accepted 7 May 2007. published online 14 June 2007.

Abstract

TIMP-1 is a promising new candidate as a prognostic marker in colorectal and breast cancer. We now describe the discovery of two alternatively spliced variants of TIMP-1 mRNA. The two variants lacking exon 2 (del-2) and 5 (del-5), respectively, were identified in human cancer cell lines by RT-PCR. The del-2 variant was, furthermore, detected in extracts from 12 colorectal cancer tissue samples. By western blotting additional bands of lower molecular mass than full-length TIMP-1 were identified in tumor tissue, but not in plasma samples obtained from cancer patients.

The two splice variants of TIMP-1 may hold important clinical information, and either alone or in combination with measurement of full-length TIMP-1 they may improve the prognostic and/or predictive value of TIMP-1 analyses.

Keywords: TIMP-1, Splice variants, Colon cancer, Cancer cell lines

Abbreviations: CRC, colorectal cancer, ELISA, enzyme-linked immunosorbent assay, GAPDH, glyceraldehyde-3-phosphate dehydrogenase, HRP, horseradish peroxidase, MMP, matrix metalloproteinase, α-SMA, α-smooth muscle actin, TIMP-1, tissue inhibitor of metalloproteinases-1, TNP, trinitrophenyl hapten