Proteomic profiling identifies afamin as a potential biomarker for ovarian cancer.
OBJECTIVE
To discover and validate serum glycoprotein biomarkers in ovarian cancer using proteomic-based approaches.


METHODS
Serum samples from a "discovery set" of 20 patients with ovarian cancer or benign ovarian cysts or healthy volunteers were compared by fluorescence two-dimensional differential in-gel electrophoresis and parallel lectin-based two-dimensional profiling. Validation of a candidate biomarker was carried out with Western blotting and immunoassay (n = 424).


RESULTS
Twenty-six proteins that changed significantly were identified by mass spectrometric sequencing. One of these, confirmed by Western blotting, was afamin, a vitamin E binding protein, with two isoforms decreasing in patients with ovarian cancer. Validation using cross-sectional samples from 303 individuals (healthy controls and patients with benign, borderline, or malignant ovarian conditions and other cancers) assayed by ELISA showed significantly decreased total afamin concentrations in patients with ovarian cancer compared with healthy controls (P = 0.002) and patients with benign disease (P = 0.046). However, the receiver operating characteristic areas for total afamin for the comparison of ovarian cancer with healthy controls or benign controls were only 0.67 and 0.60, respectively, with comparable figures for CA-125 being 0.92 and 0.88 although corresponding figures for a subgroup of samples analyzed by isoelectric focusing for afamin isoform 2 were 0.85 and 0.79. Analysis of a further 121 samples collected prospectively from 9 patients pretreatment through to relapse indicated complementarity of afamin with CA-125, including two cases in whom CA-125 was noninformative.


CONCLUSIONS
Afamin shows potential complementarity with CA-125 in longitudinal monitoring of patients with ovarian cancer, justifying prospective larger-scale investigation. Changes in specific isoforms may provide further information.
Jackson
David
D
Cancer Research UK Clinical Centre, St James's University Hospital, Leeds, United Kingdom.
Craven
Rachel A
RA
Hutson
Richard C
RC
Graze
Ina
I
Lueth
Paul
P
Tonge
Robert P
RP
Hartley
Joanne L
JL
Nickson
Janice A
JA
Rayner
Steve J
SJ
Johnston
Colin
C
Dieplinger
Benjamin
B
Hubalek
Michael
M
Wilkinson
Nafisa
N
Perren
Timothy J
TJ
Kehoe
Sean
S
Hall
Geoffrey D
GD
Daxenbichler
Guenter
G
Dieplinger
Hans
H
Selby
Peter J
PJ
Banks
Rosamonde E
RE
D014408:Biomarkers, Tumor / Q000097:blood*; D015153:Blotting, Western; D018394:CA-125 Antigen / Q000097:blood; D002352:Carrier Proteins / Q000097:blood*; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D010051:Ovarian Neoplasms / Q000097:blood* / Q000235:genetics; D020033:Protein Isoforms / Q000097:blood; D040901:Proteomics; D012372:ROC Curve; D012709:Serum Albumin; D000075462:Serum Albumin, Human
C088246:AFM protein, human; D014408:Biomarkers, Tumor; D018394:CA-125 Antigen; D002352:Carrier Proteins; D006023:Glycoproteins; D020033:Protein Isoforms; D012709:Serum Albumin; D000075462:Serum Albumin, Human