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Assessment of ApoC1, LuzP6, C12orf75 and OCC-1 in cystic glioblastoma using MALDI–TOF mass spectrometry, immunohistochemistry and qRT-PCR
- Source :
- Medical Molecular Morphology
- Publication Year :
- 2019
- Publisher :
- Springer Japan, 2019.
-
Abstract
- 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% (LuzP61–29) and 96% (LuzP630–58) 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. Electronic supplementary material The online version of this article (10.1007/s00795-019-00223-8) contains supplementary material, which is available to authorized users.
- Subjects :
- 0301 basic medicine
Cystic glioblastoma
Male
Malignant glioma
OCC-1
Pathology and Forensic Medicine
03 medical and health sciences
0302 clinical medicine
ApoC1
Protein purification
medicine
Humans
Cyst
Molecular Biology
Messenger RNA
Original Paper
Apolipoprotein C-I
biology
Chemistry
Reverse Transcriptase Polymerase Chain Reaction
General Medicine
Middle Aged
medicine.disease
Molecular biology
Molecular medicine
Immunohistochemistry
Neoplasm Proteins
030104 developmental biology
Real-time polymerase chain reaction
C12orf75
030220 oncology & carcinogenesis
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
biology.protein
Female
LuzP6
Antibody
Glioblastoma
MALDI–TOF
Subjects
Details
- Language :
- English
- ISSN :
- 18601499 and 18601480
- Volume :
- 52
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Medical Molecular Morphology
- Accession number :
- edsair.doi.dedup.....0e62d137e5b8cc384fb7c7a013fb518d