Tissue microarray analysis of eIF4E and its downstream effector proteins in human breast cancer

人类乳腺癌中 eIF4E 及其下游效应蛋白的组织微阵列分析

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作者:Heather E Kleiner, Prasad Krishnan, Jesse Tubbs, Mark Smith, Carol Meschonat, Runhua Shi, Mary Lowery-Nordberg, Patrick Adegboyega, Marcia Unger, James Cardelli, Quyen Chu, J Michael Mathis, John Clifford, Arrigo De Benedetti, Benjamin D L Li

Background

Eukaryotic initiation factor 4E (eIF4E) is elevated in many cancers and is a prognostic indicator in breast cancer. Many pro-tumorigenic proteins are selectively translated via eIF4E, including c-Myc, cyclin D1, ornithine decarboxylase (ODC), vascular endothelial growth factor (VEGF) and Tousled-like kinase 1B (TLK1B). However, western blot analysis of these factors in human breast cancer has been limited by the availability of fresh frozen tissue and the labor-intensive nature of the multiple assays required. Our goal was to validate whether formalin-fixed, paraffin-embedded tissues arranged in a tissue microarray (TMA) format would be more efficient than the use of fresh-frozen tissue and western blot to test multiple downstream gene products.

Conclusion

We conclude that the TMA technique yields similar results as the western blot technique and can be more efficient and thorough in the evaluation of several products downstream of eIF4E.

Results

Breast tumor TMAs were stained immunohistochemically and quantitated using the ARIOL imaging system. In the TMAs, eIF4E levels correlated strongly with c-Myc, cyclin D1, TLK1B, VEGF, and ODC. Western blot comparisons of eIF4E vs. TLK1B were consistent with the immunohistochemical results. Consistent with our previous western blot results, eIF4E did not correlate with node status, ER, PR, or HER-2/neu.

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