Identification of protein marker in vaginal wall tissues of women with stress urinary incontinence by protein chip array.
Identification of protein marker in vaginal wall tissues of women with stress urinary incontinence by protein chip array.
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DOI：
10.1111/j.1447-0756.2011.01690.x
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发表时间：
2012-01
期刊：
影响因子：
--
通讯作者：
Chen B
中科院分区：
文献类型：
--
作者：
Wen Y;Whitin J;Yu T;Cohen H;Polan ML;Chen B
我们试图调查蛋白质生物标志物的压力性尿失禁（SUI）在阴道组织中使用表面增强激光解吸/电离飞行时间质谱（SELDI-TOF MS），并检查这是否是一个可靠的方法来检查蛋白质在小组织标本。我们比较了SUI患者和对照组阴道组织的蛋白质表达谱。随后通过弱阳离子交换、反相分离、凝胶电泳、胰蛋白酶消化鉴定22.6 kDa峰，然后通过基质辅助激光解吸/电离质谱（MALDI MS）和MALDI MS-MS进行分析。通过Western印迹和免疫组织化学确认生物标志物身份和表达水平。与对照组相比，SUI患者中22.6 kDa蛋白（SM-22α）的表达显著升高。3 x 3 mm的组织样本足以进行识别。Western blot/免疫组化证实了SELDI-TOS MS结果。SM-22α是肌成纤维细胞的标志物，被鉴定为SUI的生物标志物。SELDI-TOF MS的差异蛋白质谱是泌尿妇科研究的一个强大而可靠的工具，因为它允许我们使用小的组织样本同时研究一系列蛋白质。
We sought to investigate protein biomarkers for stress urinary incontinence (SUI) in vaginal tissues using Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and examine if this is a reliable methodology to examine proteins in small tissue specimens. We compared protein expression profile of vaginal tissue from women with SUI and continent controls. A 22.6 kDa peak was identified by subsequent weak cation-exchange, reverse-phase fractionation, gel electrophoresis, trypsin digestion, then analyzed by matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) and MALDI MS-MS. Biomarker identity and expression level were confirmed by Western-blotting and immunohistochemistry. Expression of the 22.6 kDa protein, identified as SM-22α, was significantly higher in women with SUI versus controls. A 3×3mm tissue sample was sufficient for identification. Western blot/immunohistochemistry confirmed the SELDI-TOS MS findings. SM-22α, a marker for myofibroblasts, was identified as a biomarker of SUI. Differential protein profiling by SELDI-TOF MS is a powerful and reliable tool for urogynecological research since it allows us to study an array of proteins simultaneously using small tissue samples.
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