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Identification of genes differentially expressed between benign and osteopontin transformed rat mammary epithelial cells

Vittal V Kurisetty1, Patrick G Johnston1, Philip S Rudland2 and Mohamed K El-Tanani1*

Author Affiliations

1 Centre for Cancer Research and Cell Biology (CCRCB), Queen's University Belfast, Belfast, BT9 7BL, UK

2 Cancer and Polio Research Fund Laboratories, School of Biological Sciences, University of Liverpool, PO. Box 147, Liverpool, L69 7ZB, UK

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BMC Research Notes 2009, 2:15  doi:10.1186/1756-0500-2-15

Published: 3 February 2009



Osteopontin is a secreted, integrin-binding and phosphorylated acidic glycoprotein which has an important role in tumor progression.


In this study, we have utilized suppressive subtractive hybridization (SSH) to evaluate OPN regulated gene expression, using the Rama 37 benign non-invasive rat mammary cell line and a subclone, Rama 37-OPN. Rama 37-OPN was produced by stably transfecting Rama 37 with an OPN expression vector and it demonstrates increased malignant properties in vitro. Sequence and expression array analysis of the respective cDNA libraries of over 1600 subtracted cDNA fragments revealed 982 ESTs, 45 novel sequences and 659 known genes. The known up-regulated genes in the Rama 37-OPN library code for proteins with a variety of functions including those involved in metabolism, cell adhesion and migration, signal transduction and in apoptosis. Four of the most differentially expressed genes between the benign and in vitro malignant rat mammary cell lines are tumor protein translationally controlled I (TPTI), aryl hydrocarbon receptor nuclear translocator (ARNT), ataxia telangiectasia mutated (ATM) and RAN GTPase (RAN). The largest difference (ca 10,000 fold) between the less aggressively (MCF-7, ZR-75) and more aggressively malignant (MDA MB 231, MDA MB 435S) human breast cancer cell lines is that due to RAN, the next is that due to osteopontin itself.


The results suggest that enhanced properties associated with the malignant state in vitro induced by osteopontin may be due to, in part, overexpression of RAN GTPase and these biological results are the subject of a subsequent publication [1].