Quantitative iTRAQ LC-MS/MS Proteomics Reveals Transcription Factor Crosstalk and Regulatory Networks in Hypopharyngeal Squamous Cell Carcinoma

Abstract

To date, no effective therapeutic treatments have been developed for hypopharyngeal squamous cell carcinoma (HPSCC), a disease that has a five-year survival rate of approximately 31% because of its late diagnosis and aggressive nature. Despite recent improvements in diagnostic methods, there are no effective measures to prevent or detect HPSCC in an early stage. The goal of the current study was to identify molecular biomarkers and networks that can facilitate the speedy identification of HPSCC patients who could benefit from individualized treatment. Isobaric tags for relative and absolute quantification (iTRAQ) labeling was employed with two-dimensional liquid chromatography-tandem mass spectrometry to identify quantitatively the differentially expressed proteins among three types of HPSCC disease stages. The iTRAQ results were evaluated by literature searches and western blot analysis. For example, FUBP1, one of 412 proteins with significantly altered expression profiles, was confirmed to have elevated expression in fresh HPSCC tissues. Integrin-mediated cell matrix adhesion and actin filament-inducing cytoskeleton remodeling were the cellular events that were the most relevant to HPSCC tumorigenesis and the metastatic process. The construction of transcriptional regulation networks led to the identification of key transcriptional regulators of tumor development and lymph node metastasis of HPSCC, including Sp1, c-Myc and p53. Additionally, our study indicated that the interactions among Sp1, c-Myc and p53 may play vital roles in the carcinogenesis and metastasis of HPSCC.

Metadherin Regulation of Vascular Endothelial Growth Factor Expression Is Dependent Upon the PI3K/Akt Pathway in Squamous Cell Carcinoma of the Head and Neck

“…All of the western blotting analyses were performed as described previously. 34 In brief, 30 μg of protein was separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto a Polyvinalidene Fluoride membrane (Millipore, Bedford, MA). The blotted membranes were incubated with primary antibodies against MTDH (1:800; Proteintech Group, Inc, Chicago, IL), VEGF (1:1000; Santa Cruz, CA), pan-Akt (1:1000; Cell Signaling Technology, Inc, Beverly, MA), phospho-Akt (Thr308) (1:800; Cell Signaling Technology, Inc), and then with the corresponding secondary antibody (1:3000; Beyotime, Shanghai, China).…”

Interaction between TNFR1 and TNFR2 dominates the clinicopathologic features of human hypopharyneal carcinoma

“…From oncologic aspects, HPSCC possesses the nature of untamable malignant progression and therapeutic resistance, including aggressive infiltration and invasion, early lymph node metastasis, and insensitivity to radiotherapy and/or chemotherapy, with resultant relatively high recurrence and mortality rates [3][4][5]. Up till present, many important factors responsible for regulating cell proliferation and apoptosis have been associated with the biological behaviors of HPSCC [6,7]. TNFR1 and TNFR2 are two membranous receptors that anchor on the surface of cell membrane, both of which belong to the TNF receptor superfamily [8,9].…”

Identification of Transcription Factor YY1 as a Regulator of a Prostate Cancer-Specific Pathway Using Proteomic Analysis

“…Proteomics is a powerful tool that can be used to assess various transcriptional regulatory mechanisms and direct interactions between transcription factors and proteins 35 - 38 . We found that many of the proteins that were over-expressed in prostate cancer cell lines, compared to a non-cancerous prostate cell line, are involved in mitochondrial energy metabolism.…”