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 language | English (US) |
|---|---|
| Pages (from-to) | 502-506 |
| Number of pages | 5 |
| Journal | Proteomics - Clinical Applications |
| Volume | 6 |
| Issue number | 9-10 |
| DOIs | |
| State | Published - Oct 2012 |
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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 journal › Article
}
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 -