Abstract
Triple negative breast cancer (TNBC) is a heterogeneous disease with more aggressive clinical courses than other subtypes of breast cancer. In this study, we performed high-resolution mass spectrometry-based quantitative proteomics with TNBC clinical tissue specimens to explore the early and sensitive diagnostic signatures and potential therapeutic targets for TNBC patients. We performed an iTRAQ labeling coupled LC-MS/MS approach to explore the global proteome in tumor tissues and corresponding para-tumor tissues from 24 patients with grade I-II and grade III primary TNBC. Relative peptide quantification and protein identification were performed by Proteome Discoverer™ software with Mascot search engine. Differentially expressed proteins were analyzed by bioinformatic analyses, including GO function classification annotation and KEGG enrichment analysis. Pathway analyses for protein-protein interactions and upstream regulations of differentially expressed candidates were performed by Ingenuity Pathway Analysis (IPA) software. Totally, 5401 unique proteins were identified and quantified in different stage of TNBCs. 845 proteins were changed in patients with grade I or II TNBC, among which 304 were up-regulated and 541 were down-regulated. Meanwhile, for patients with grade III TNBC, 358 proteins were increased and 651 proteins were decreased. Comparing to para-cancerous tissues, various signaling pathways and metabolic processes, including PPAR pathways, PI3K-Akt pathway, one-carbon metabolism, amino acid synthesis, and lipid metabolism were activated in TNBC cancer tissues. Death receptor signaling was significantly activated in grade I-II TNBCs, however, remarkably inhibited in grade III TNBCs. Western blot experiments were conducted to validate expression levels of CYCS, HMGA1 and XIAP with samples from individual patients. Overall, our proteomic data presented precise quantification of potential signatures, signaling pathways, regulatory networks, and characteristic differences in each clinicopathological subgroup. The proteome provides complementary information for TNBC accurate subtype classification and therapeutic targets research.
Highlights
MethodsWe performed an iTRAQ-labeling coupled LC-MS/MS approach to explore the global proteome in tumor tissues and corresponding para-tumor tissues from 24 patients with grade I-II and grade III primary Triple negative breast cancer (TNBC)
Triple negative breast cancer (TNBC) is a heterogeneous disease with more aggressive clinical courses than other subtypes of breast cancer
5,401 unique proteins were identified and quantified in different stage of TNBCs. 865 proteins were changed in patients with grade I or II TNBC, among which 309 were up-regulated and 556 were down-regulated
Summary
We performed an iTRAQ-labeling coupled LC-MS/MS approach to explore the global proteome in tumor tissues and corresponding para-tumor tissues from 24 patients with grade I-II and grade III primary TNBC. This study and consent procedure was approved by the Ethics Committee of Fujian Medical University Union Hospital (Fujian, China). The paired TNBC samples used in this study were obtained from patients who underwent breast surgery (Total mastectomy or breast conserving surgery) from the department of breast surgery, Fujian Medical. Resected primary tumor tissues and paired non-tumor breast tissues were selected from 12 patients with grade I-II as well as 12 patients with grade III. All grade scoring were performed by two independent pathologists without knowledge of patients status. Clinical information was collected, including gender, age, histological grade, Ki-67 labeling index, tumor number, diameter of tumor, status of axillary lymph node metastasis. The tissue samples for the proteomic analysis were frozen in liquid nitrogen for storage before use
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