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Proteomics Clin Appl. 2012 Oct;6(9-10):502-6. doi: 10.1002/prca.201200018.

A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue.

Author information

1
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.

Abstract

PURPOSE:

Quantitation of β-tubulin isotype expression in taxane resistant human tumor tissue has been difficult to achieve because of the limited availability of validated antibodies. Here we present a label-free MS method to quantitate relative expression levels of β-tubulin isotypes.

EXPERIMENTAL DESIGN:

Using isotype-specific reporter peptides, we determined relative β-tubulin isotype expression levels in human lung tumor tissue.

RESULTS:

Four reporter peptides were chosen to quantitate the βI/βII, βIV, βIII, and βV tubulin isotypes. These peptides were validated using human cancer cell lines. The label-free method was then used to determine β-tubulin isotype expression in nine human lung tumor samples, which had been described as high or low βIII-tubulin expressing using immunohistochemistry. It was found that βI/βII (accounting for 18.7-65.7% of total β-tubulin) and βIVa/βIVb (26.3-79.1%) were the most abundant isotypes and that the βIII (0-8.9%) and βV (1.0-10.4%) were less abundant in the tissue. We also categorized the samples as high or low βIII-tubulin expressing.

CONCLUSION AND CLINICAL RELEVANCE:

With this method we can determine the relative expression levels of β-tubulin isotypes in human tumor tissue. This method will facilitate studies assessing the use of tubulin isotypes as biomarkers of taxane resistance.

PMID:
22996942
PMCID:
PMC3484181
DOI:
10.1002/prca.201200018
[Indexed for MEDLINE]
Free PMC Article

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