A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue.
A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue.
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DOI：
10.1002/prca.201200018
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发表时间：
2012-10
影响因子：
2
通讯作者：
Angeletti, Ruth Hogue
中科院分区：
文献类型：
--
作者：
Miller, Leah M.;Yang, Chia-Ping Huang;Xiao, Hui;Isaac, Sylvie;Seve, Pascal;Dumontet, Charles;Horwitz, Susan Band;Angeletti, Ruth Hogue
关键词：
由于有效抗体的可用性有限，对紫杉烷抗性人类肿瘤组织中 β-微管蛋白同种型表达的定量一直难以实现。在这里，我们提出了一种无标记质谱方法来定量 β-微管蛋白同种型的相对表达水平。使用同种型特异性报告肽，我们确定了人肺肿瘤组织中的相对 β-微管蛋白同种型表达水平。选择四种报告肽来定量 βI/βII、βIV、βIII 和 βV 微管蛋白同种型。这些肽使用人类癌细胞系进行了验证。然后使用无标记方法测定九个人肺肿瘤样本中的β-微管蛋白同种型表达，使用免疫组织化学将其描述为高或低βIII-微管蛋白表达。研究发现组织中βI/βII（占总β微管蛋白的18.7%~65.7%）和βIVa/βIVb（26.3%~79.1%）是最丰富的同种型，而βIII（0~8.9%）和βV（1.0~10.4%）的同种型较少。我们还将样品分为高或低 βIII-微管蛋白表达。通过这种方法，我们可以测定人类肿瘤组织中β-微管蛋白同种型的相对表达水平。该方法将有助于评估微管蛋白同种型作为紫杉烷抗性生物标志物的使用的研究。
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 mass spectrometry method to quantitate relative expression levels of β–tubulin isotypes. Using isotype specific reporter peptides, we determined relative β–tubulin isotype expression levels in human lung tumor tissue. 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. 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.
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影响因子：
64.8
作者：
SCHIFF, PB;FANT, J;HORWITZ, SB
通讯作者：
HORWITZ, SB
影响因子：
2.9
作者：
Verdier-Pinard, P;Shahabi, S;Horwitz, SB
通讯作者：
Horwitz, SB
影响因子：
2.9
作者：
Leandro-Garcia, Luis J.;Leskelae, Susanna;Rodriguez-Antona, Cristina
通讯作者：
Rodriguez-Antona, Cristina
影响因子：
2.9
作者：
DERRY, WB;WILSON, L;JORDAN, MA
通讯作者：
JORDAN, MA
影响因子：
2.9
作者：
Winefield RD;Williams TD;Himes RH
通讯作者：
Himes RH