Abstract
The purpose of this study was to develop quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the analysis of proteins involved in metastasis of breast cancer for diagnosis and determining disease prognosis, as well as to further our understand of metastatic mechanisms. We have previously demonstrated that the protein type XIV collagen may be specifically expressed in metastatic tissues by two dimensional LC-MS/MS. In this study, we developed quantitative LC-MS/MS methods for type XIV collagen. Type XIV collagen was quantified by analyzing 2 peptides generated by digesting type XIV collagen using stable isotope-labeled peptides. The individual concentrations were equivalent between 2 different peptides of type XIV collagen by evaluation of imprecise transitions and using the best transition for the peptide concentration. The results indicated that type XIV collagen is highly expressed in metastatic tissues of patients with massive lymph node involvement compared to non-metastatic tissues. These findings were validated by quantitative real-time RT-PCR. Further studies on type XIV collagen are desired to verify its role as a prognostic factor and diagnosis marker for metastasis.
Highlights
The 5-year breast cancer survival rate is generally greater than 90% but it is roughly 55% in women with numerous lymph node metastases [1,2,3]
By comprehensive analysis using 2D LC-MS/MS, 34 proteins were screened as specific candidate proteins expressed highly in massive lymph node metastasis breast cancer [5]
Type XIV collagen may not be an appropriate target of antibody drugs because it is present in normal extracellular matrix, it may be a potential biomarker of metastasis and useful in evaluation of patient prognosis
Summary
The 5-year breast cancer survival rate is generally greater than 90% but it is roughly 55% in women with numerous lymph node metastases [1,2,3]. The prognosis of breast cancer associated with nodal metastasis, age, tumor size (pT), histological grade of tumor, estrogen and progesterone receptors status, human epidermal growth factor receptors (HERs) status, and breast cancer (BRCA) gene mutations. Nodal status (including number and location of nodes) correlates with disease-free and overall survival better than any other prognostic factor. As metastasized and recurrent breast cancers (especially breast cancer with massive nodal metastasis) are difficult to cure, the development of new therapeutic agents including molecular targeted drugs is awaited. We have searched for the role of lymph node- and metastasis-related biomarkers [4].
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