Evangelou Petros, Groll Mathias, Oppermann Henry, Gaunitz Frank, Eisenlöffel Christian, Müller Wolf, Eschrich Klaus, Schänzer Anne, Nestler Ulf
Department of Neurosurgery, University Hospital Leipzig, Liebigstrasse 20, 04103, Leipzig, Germany.
Institute of Neuropathology, University Hospital Leipzig, Liebigstrasse 26, 04103, Leipzig, Germany.
Med Mol Morphol. 2019 Dec;52(4):217-225. doi: 10.1007/s00795-019-00223-8. Epub 2019 Apr 20.
Mass spectrometric analysis of glioblastoma cyst fluids has disclosed a protein peak with m/z 6424-6433. Among the proteins, potentially generating this peak are ApoC1 and LuzP6. To further elucidate protein expression of glioblastoma cells, we analyzed MALDI-TOF results of cyst fluid, performed immunohistochemistry and mRNA analysis. MALDI-TOF protein extraction from 24 glioblastoma cyst fluids was performed with a weak cation exchange. 50 glioblastoma samples were stained with two custom-made antibodies against LuzP6 and commercial antibodies against ApoC1, C12orf75 and OCC-1 and analyzed. For mRNA detection, 16 tissue samples were stored in RNAlater, extracted using the miRNeasy kit and reversely transcribed. For 12 patients, synopsis of results from all three examinations was possible. MALDI-TOF confirmed the peak at 6433 Da in 75% of samples. Immunohistochemically, LuzP6 was detected in 92% (LuzP6) and 96% (LuzP6) of samples and ApoC1 in 66%. Mean mRNA levels were highest for ApoC1, followed by LuzP6. No correlation between mRNA expression, immunohistochemical staining and intensity of the MALDI-TOF peaks was found. An unequivocal identification of one protein as the source for the 6433 peak is not possible, but our results point to ApoC1 and LuzP6 as the underlying proteins.
胶质母细胞瘤囊液的质谱分析揭示了一个质荷比为6424 - 6433的蛋白质峰。在这些蛋白质中，可能产生此峰的是载脂蛋白C1（ApoC1）和LuzP6。为了进一步阐明胶质母细胞瘤细胞的蛋白质表达情况，我们分析了囊液的基质辅助激光解吸电离飞行时间质谱（MALDI - TOF）结果，进行了免疫组织化学和mRNA分析。采用弱阳离子交换法从24份胶质母细胞瘤囊液中提取MALDI - TOF蛋白质。用两种针对LuzP6的定制抗体以及针对ApoC1、C12orf75和OCC - 1的商业抗体对50份胶质母细胞瘤样本进行染色并分析。对于mRNA检测，将16份组织样本保存在RNA Later中，使用miRNeasy试剂盒提取并进行逆转录。对于12名患者，可以综合这三项检查的结果。MALDI - TOF在75%的样本中确认了6433 Da处的峰。免疫组织化学检测发现，92%（LuzP6）和96%（LuzP6）的样本中检测到LuzP6，66%的样本中检测到ApoC1。平均mRNA水平以ApoC1最高，其次是LuzP6。未发现mRNA表达、免疫组织化学染色与MALDI - TOF峰强度之间存在相关性。虽然无法明确鉴定出一种蛋白质作为6433峰的来源，但我们的结果表明ApoC1和LuzP6是潜在的相关蛋白质。
