应用基质辅助激光解吸附电离飞行时间质谱分析宫颈鳞癌早期浸润蛋白质

doi:10.3877/cma.j.issn.1673-5250.2011.04.011

目的

探讨基质辅助激光解吸附电离飞行时间质谱(matrix assisted laser desorption ionization time of flight mass spectrometry，MALDI-TOF-MS)对宫颈鳞癌(squamous carcinoma of the cervix，SCC)组织切片直接分析，获取宫颈鳞癌早期浸润的分子标志物的有效性。

方法

利用超微量点样枪将化学基质直接点样到宫颈鳞癌组织切片上，然后利用基质辅助激光解吸附电离飞行时间质谱直接扫描宫颈鳞癌组织切片，分析宫颈鳞癌组织蛋白质分子。

结果

获得的25种蛋白质质谱峰强度比较，差异有显著意义(P<0.05)。其中7种差异蛋白质有分类意义，其质/核(M/Z)分别为：m/z 3 433，m/z 4786，m/z 10 092, m/z 10 835，m/z 15 114，m/z 15 854和m/z 16 345。差异蛋白质m/z 3 433，m/z 15 114，m/z 15 854在宫颈鳞癌鳞状上皮组织高度表达，可作为宫颈鳞癌早期诊断的标志蛋白；差异蛋白质m/z 16 345在宫颈鳞癌的间质组织高度表达，可作为宫颈鳞癌早期间质浸润和扩散诊断的标志蛋白。

结论

利用基质辅助激光解吸附电离飞行时间质谱技术成功获取一组差异蛋白质。该差异蛋白组可望作为宫颈鳞癌早期诊断的标志蛋白。

关键词: 宫颈鳞癌, 早期浸润, 差异蛋白质组, 基质辅助激光解吸附电离飞行时间质谱

Objective

To analyze the distribution of proteins in cervical invasive cancer which were profiled directly by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

Methods

A saturated sinapinic acid (SA) solution was prepared at the concentration of 35 mg/mL in 50: 50 acetonitrile/0.1% TFA in water. Matrix sinapinic acid solution was deposited onto tissue sections by using a low-volume pipette. Samples were analyzed directly by matrix assisted laser desorption ionization time of flight mass spectrometry.

Results

A total of 25 types of candidate proteins were screened, and 7 candidate proteins were significant (m/z 3 433, m/z 4786, m/z 10 092, m/z 10 835, m/z 15 114, m/z 15 854, m/z 16 345). Importantly, candidate proteins m/z 3 433, m/z 15 114 and m/z 15 854 were expressed overly in the epithelium of cervical cancer, whereas candidate protein m/z 16 345 was expressed highly in the stromal extracellular matrix (ECM) of cervical cancer.

Conclusions

The differences of expressions and distributions of candidate proteins were found in the epithelium between cervical cancer and normal control group, suggesting that these candidate proteins may be as now biomarkers to be used in improving the predictive value of cervical invasive cancer.

Key words: squamous carcinoma of the cervix (SCC), early infiltrate, proteomic difference, matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS)

图1　宫颈鳞癌组织切片

图2　化学基质点阵

图3　正常宫颈组织蛋白质质谱图(Intens. [a.u.])

图4　宫颈鳞癌早期浸润组织蛋白质谱图(Intens. [a.u.])

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Direct Profiling of Proteins Distribution in Cervical Invasive Cancer by Matrix Assisted Laser Desorption Ionization Mass Spectrometry

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Direct Profiling of Proteins Distribution in Cervical Invasive Cancer by Matrix Assisted Laser Desorption Ionization Mass Spectrometry