Wen Yan, Whitin John, Yu Tom, Cohen Harvey, Polan Mary Lake, Chen Bertha
Department of Obstetrics and Gynecology Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
J Obstet Gynaecol Res. 2012 Jan;38(1):89-96. doi: 10.1111/j.1447-0756.2011.01690.x. Epub 2011 Dec 5.
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 22.6kDa peak was identified by subsequent weak cation-exchange, reverse-phase fractionation, gel electrophoresis, and 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 22.6kDa protein, identified as SM-22α, was significantly higher in women with SUI versus controls. A 3×3-mm 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 as it allows us to study an array of proteins simultaneously using small tissue samples.
我们试图利用表面增强激光解吸/电离飞行时间质谱(SELDI-TOF MS)研究阴道组织中压力性尿失禁(SUI)的蛋白质生物标志物,并检验这是否是一种检测小组织标本中蛋白质的可靠方法。
我们比较了SUI女性和控尿对照者阴道组织的蛋白质表达谱。通过后续的弱阳离子交换、反相分级分离、凝胶电泳和胰蛋白酶消化,鉴定出一个22.6kDa的峰,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。通过蛋白质印迹法和免疫组织化学法确认生物标志物的身份和表达水平。
被鉴定为SM-22α的22.6kDa蛋白质在SUI女性中的表达明显高于对照组。3×3毫米的组织样本足以进行鉴定。蛋白质印迹法/免疫组织化学法证实了SELDI-TOS MS的结果。
SM-22α,一种肌成纤维细胞标志物,被鉴定为SUI的生物标志物。SELDI-TOF MS进行的差异蛋白质谱分析是泌尿妇科研究的一种强大而可靠的工具,因为它使我们能够使用小组织样本同时研究一系列蛋白质。