• Breast cancer;
  • ITIH4;
  • LC-MS/MS;
  • Quantification;
  • Serum


Purpose: Various studies exploring the potential of the low-molecular-weight serum peptidome have identified proteolytic cleavage products of inter-α-trypsin inhibitor heavy chain-4 (ITIH4) as potential markers for different types of cancer, presumably generated by tumor-associated exoproteases. However, further elucidation of the discriminative properties of such peptides requires specific quantitative analytical methods.

Experimental design: Using a recently developed and fully validated liquid chromatography–tandem mass spectrometric method, we have compared absolute serum concentrations of eight peptides derived from ITIH4 [658–687] to [667–687] (ITIH4-30 to −21) between breast cancer patients (n=45) and controls (n=78). Furthermore, serum samples obtained before and after surgical removal of the tumor were analyzed (n=30).

Results: The inter-individual variability in measured serum concentrations was high. Nevertheless, most peptides showed a tendency toward elevated levels in the presence of the breast cancer tumor. Significantly increased serum concentrations were observed in the breast cancer group for ITIH4-25 (p=0.036) and −29 (p=0.015). Intra-individual comparisons of serum obtained before and after surgery showed significantly decreased serum levels after surgery for seven of the ITIH4-derived peptides (p<0.02).

Conclusions and clinical relevance: The obtained results particularly suggest potential for these ITIH4-derived peptides in the follow-up of breast cancer after surgery.