Abstract:
Objective The objective of this study is to detect ovarian cancer tumor markers using proteomics approaches of isobaric tags for relative and absolute quantification (iTRAQ) and two-dimensional nano high-performance liquid chromatography-electrospray-OrbiTrap mass spectrometry (2D nano HPLC-ESI-OrbiTrap MS/MS) to improve the early diagnosis rate.
Methods Serum samples from 10 patients with ovarian cancer, 10 with benign ovarian tumor, and 10 normal women subjects were pooled with equal volume. A high abundance of protein was depleted after the balanced mix of the samples. The samples were then detected and identified by the iTRAQ– coupled 2D nano HPLC-ESI-OrbiTrap MS/MS technology. The proteins with significant differences were checked using Western blot in the 56 serum samples.
Results A total of 326 proteins were identified. Among these proteins, eight important proteins were interpreted, including proteins PARD3, SRP1-alpha, LAMA4, LRP16, IgSF2, NRAMP1, NF-E2-related factor 1, and APOA4. Using Western blot technology, three important proteins with significant differences were validated, namely, proteins APOA4, PARD3, and NRAMP1.
Conclusion The eight important proteins discovered using iTRAQ may be the potential tumor makers of ovarian cancer. As a novel protein, APOA4 may become a biological marker to differentiate malignant from benign tumor in a serum that has an elevated CA-125.
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