Feasibility study on measuring selected proteins in malignant melanoma tissue by SRM quantification.
Currently there are no clinically recognized molecular biomarkers for malignant melanoma (MM) for either diagnosing disease stage or measuring response to therapy. The aim of this feasibility study was to develop targeted selected reaction monitoring (SRM) assays for identifying candidate protein biomarkers in metastatic melanoma tissue lysate. In a pilot study applying the SRM assay, the tissue expression of nine selected proteins [complement 3 (C3), T-cell surface glycoprotein CD3 epsilon chain E (CD3E), dermatopontin, minichromosome maintenance complex component (MCM4), premelanosome protein (PMEL), S100 calcium binding protein A8 (S100A8), S100 calcium binding protein A13 (S100A13), transgelin-2 and S100B] was quantified in a small cohort of metastatic malignant melanoma patients. The SRM assay was developed using a TSQ Vantage triple quadrupole mass spectrometer that generated highly accurate peptide quantification. Repeated injection of internal standards spiked into matrix showed relative standard deviation (RSD) from 6% to 15%. All nine target proteins were identified in tumor lysate digests spiked with heavy peptide standards. The multiplex SRM peptide assay panel was then measured and quantified on a set of frozen MM tissue samples obtained from the Malignant Melanoma Biobank collected in Lund, Sweden. All nine proteins could be accurately quantified using the new SRM assay format. This study provides preliminary data on the heterogeneity of biomarker expression within MM patients. The S100B protein, which is clinically used as the pathology identifier of MM, was identified in 9 out of 10 MM tissue lysates. The use of the targeted SRM assay provides potential advancements in the diagnosis of MM that can aid in future assessments of disease in melanoma patients.
Departments of †Oncology, ∥Surgery, and ⊥Cancer Epidemiology, Clinical Sciences, and ‡Centre of Excellence in Biological and Medical Mass Spectrometry, Lund University , 221 85 Lund, Sweden.
Charlotte
C
Welinder
Göran
G
Jönsson
Christian
C
Ingvar
Lotta
L
Lundgren
Bo
B
Baldetorp
Håkan
H
Olsson
Thomas
T
Breslin
Melinda
M
Rezeli
Bo
B
Jansson
Thomas
T
Laurell
Thomas E
TE
Fehniger
Elisabet
E
Wieslander
Krzysztof
K
Pawlowski
György
G
Marko-Varga
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D018070:Biological Specimen Banks; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D005240:Feasibility Studies; D005260:Female; D015870:Gene Expression; D006801:Humans; D008207:Lymphatic Metastasis; D008297:Male; D008545:Melanoma / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D064568:S100 Calcium Binding Protein beta Subunit / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012878:Skin Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D014020:Tissue Extracts / Q000737:chemistry
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D064568:S100 Calcium Binding Protein beta Subunit; C579160:S100B protein, human; D014020:Tissue Extracts