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首页> 外文期刊>Cell motility and the cytoskeleton >Comparison of beta-tubulin mRNA and protein levels in 12 human cancer cell lines.
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Comparison of beta-tubulin mRNA and protein levels in 12 human cancer cell lines.

机译：12种人类癌细胞系中β-微管蛋白mRNA和蛋白质水平的比较。

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Antimitotic drugs are chemotherapeutic agents that bind tubulin and microtubules. Resistance to these drugs is a major clinical problem. One hypothesis is that the cellular composition of tubulin isotypes may predict the sensitivity of a tumor to antimitotics. Reliable and sensitive methods for measuring tubulin isotype levels in cells and tissues are needed to address this hypothesis. Quantitative measurements of tubulin isotypes have frequently relied upon inferring protein amounts from mRNA levels. To determine whether this approach is justified, protein and mRNA levels of beta-tubulin isotypes from 12 human cancer cell lines were measured. This work focused on only beta-tubulin isotypes because we had readily available monoclonal antibodies for quantitative immunoblots. The percentage of beta-tubulin isotype classes I, II, III, and IVa + IVb mRNA and protein were compared. For beta-tubulin class I that comprises >50% of the beta-tubulin protein in 10 of the 12 cell lines, there was good agreement between mRNA and protein percentages. Agreement between mRNA and protein was also found for beta-tubulin class III. For beta-tubulin classes IVa + IVb, we observed higher protein levels compared to mRNA levels.Beta-tubulin class II protein was found in only four cell lines and in very low abundance. We conclude that quantitative Western blotting is a reliable method for measuring tubulin isotype levels in human cancer cell lines. Inferring protein amounts from mRNA levels should be done with caution, since the correspondence is not one-to-one for all tubulin isotypes.
机译：抗有丝分裂药物是结合微管蛋白和微管的化学治疗剂。对这些药物的耐药性是主要的临床问题。一种假设是微管蛋白同种型的细胞组成可以预测肿瘤对抗有丝分裂剂的敏感性。需要一种可靠且灵敏的方法来测量细胞和组织中微管蛋白同种型水平，以解决这一假设。微管蛋白同种型的定量测量通常依赖于从mRNA水平推断蛋白质含量。为了确定这种方法是否合理，对来自12种人类癌细胞系的β-微管蛋白同种型的蛋白质和mRNA水平进行了测量。这项工作仅集中于β-微管蛋白同种型，因为我们已经有了可用于定量免疫印迹的单克隆抗体。比较了β-微管蛋白同型I，II，III和IVa + IVb mRNA和蛋白质的百分比。对于在12种细胞系中的10种中占> 50％的β-微管蛋白的I类β-微管蛋白，mRNA和蛋白百分比之间存在良好的一致性。对于β-微管蛋白III类，也发现了mRNA和蛋白质之间的一致性。对于β-微管蛋白IVa + IVb类，我们观察到蛋白质水平高于mRNA水平.β-微管蛋白II类蛋白质仅在四个细胞系中发现并且丰度非常低。我们得出结论，定量蛋白质印迹是一种用于测量人癌细胞系中微管蛋白同种型水平的可靠方法。从mRNA水平推断蛋白质的量应谨慎，因为并非所有微管蛋白同种型都是一一对应的。

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