Proteomic analysis of osteogenic sarcoma: association of tumour necrosis factor with poor prognosis.
A significant proportion of patients with osteogenic sarcoma die from lung metastasis within 5 years of diagnosis. Molecular signatures that predict pulmonary metastasis from primary osteogenic sarcoma and identify those patients at risk would be clinically useful as prognostic markers. Protein expression profiles of two clonally related murine osteogenic sarcoma cell lines with low (K12) and high (K7M2) metastatic potential were compared using two different proteomic technologies, two-dimensional difference gel electrophoresis and cell profiling by matrix-assisted laser desorption/ionization mass spectrometry. Interrogation of a molecular pathways network database suggested several additional candidate molecules that potentially predict metastatic potential of primary osteogenic sarcoma. Two such proteins, macrophage migration inhibitory factor and tumour necrosis factor were selected for further validation studies. Western blots confirmed increased expression of both cytokines in K7M2 cells compared to K12 cells. Levels of migration inhibitory factor and tumour necrosis factor were semi-quantitatively measured in human osteogenic sarcoma samples by immunohistochemistry and were correlated with clinicopathologic parameters and patient outcomes. Multivariate survival analysis demonstrated that tumour necrosis factor expression in chemotherapy naïve osteogenic sarcoma is an independent prognostic factor for overall and metastasis-free survival. No significant differences in adverse outcomes were observed based on macrophage migration inhibitory factor expression.
Cates
Justin M M
JM
Department of Pathology, Vanderbilt University Medical Center, Nashville, TN, USA. justin.m.cates@vanderbilt.edu
Friedman
David B
DB
Seeley
Erin H
EH
Dupont
William D
WD
Schwartz
Herbert S
HS
Holt
Ginger E
GE
Caprioli
Richard M
RM
Young
Pampee P
PP
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000378:metabolism* / Q000401:mortality / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D019746:Intramolecular Oxidoreductases / Q000378:metabolism; D053208:Kaplan-Meier Estimate; D008263:Macrophage Migration-Inhibitory Factors / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D009367:Neoplasm Staging; D012516:Osteosarcoma / Q000378:metabolism* / Q000401:mortality / Q000556:secondary / Q000628:therapy; D016016:Proportional Hazards Models; D040901:Proteomics / Q000379:methods; D018570:Risk Assessment; D012307:Risk Factors; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D046888:Tissue Array Analysis; D016896:Treatment Outcome; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D055815:Young Adult
D014408:Biomarkers, Tumor; D008263:Macrophage Migration-Inhibitory Factors; D014409:Tumor Necrosis Factor-alpha; D019746:Intramolecular Oxidoreductases; C076231:Mif protein, mouse