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

Leah M. Miller, Chia-Ping H. Yang, Hui Xiao, Sylvie Isaac, Pascal Sève, Charles Dumontet, Susan Band Horwitz, Ruth Hogue Angeletti

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)502-506
Number of pages5
JournalProteomics - Clinical Applications
Volume6
Issue number9-10
DOIs
StatePublished - Oct 2012

Fingerprint

Tubulin
Mass spectrometry
Labels
Tumors
Mass Spectrometry
Tissue
Neoplasms
Peptides
Lung
Biomarkers
Design of experiments
Research Design
Immunohistochemistry
Cells
Availability
Cell Line
Antibodies

Keywords

  • β-Tubulin isotypes
  • Human lung tumors
  • Label-free quantitation
  • Mass spectrometry
  • Taxane resistance

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue. / Miller, Leah M.; Yang, Chia-Ping H.; Xiao, Hui; Isaac, Sylvie; Sève, Pascal; Dumontet, Charles; Band Horwitz, Susan; Hogue Angeletti, Ruth.

In: Proteomics - Clinical Applications, Vol. 6, No. 9-10, 10.2012, p. 502-506.

Research output: Contribution to journalArticle

Miller, Leah M. ; Yang, Chia-Ping H. ; Xiao, Hui ; Isaac, Sylvie ; Sève, Pascal ; Dumontet, Charles ; Band Horwitz, Susan ; Hogue Angeletti, Ruth. / A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue. In: Proteomics - Clinical Applications. 2012 ; Vol. 6, No. 9-10. pp. 502-506.
@article{a21977dd8d434b7c9b1b875a4e63d66a,
title = "A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue",
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.",
keywords = "β-Tubulin isotypes, Human lung tumors, Label-free quantitation, Mass spectrometry, Taxane resistance",
author = "Miller, {Leah M.} and Yang, {Chia-Ping H.} and Hui Xiao and Sylvie Isaac and Pascal S{\`e}ve and Charles Dumontet and {Band Horwitz}, Susan and {Hogue Angeletti}, Ruth",
year = "2012",
month = "10",
doi = "10.1002/prca.201200018",
language = "English (US)",
volume = "6",
pages = "502--506",
journal = "Proteomics - Clinical Applications",
issn = "1862-8346",
publisher = "Wiley-VCH Verlag",
number = "9-10",

}

TY - JOUR

T1 - A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue

AU - Miller, Leah M.

AU - Yang, Chia-Ping H.

AU - Xiao, Hui

AU - Isaac, Sylvie

AU - Sève, Pascal

AU - Dumontet, Charles

AU - Band Horwitz, Susan

AU - Hogue Angeletti, Ruth

PY - 2012/10

Y1 - 2012/10

N2 - 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.

AB - 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.

KW - β-Tubulin isotypes

KW - Human lung tumors

KW - Label-free quantitation

KW - Mass spectrometry

KW - Taxane resistance

UR - http://www.scopus.com/inward/record.url?scp=84868134195&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868134195&partnerID=8YFLogxK

U2 - 10.1002/prca.201200018

DO - 10.1002/prca.201200018

M3 - Article

C2 - 22996942

AN - SCOPUS:84868134195

VL - 6

SP - 502

EP - 506

JO - Proteomics - Clinical Applications

JF - Proteomics - Clinical Applications

SN - 1862-8346

IS - 9-10

ER -