Expression Profiles and Clinical Correlations of Degradome Components in the Tumor Microenvironment of Head and Neck Squamous Cell Carcinoma

Abstract

Purpose: Head and neck squamous cell carcinomas (HNSCC) are characterized by high morbidity and mortality, largely due to the high invasive and metastatic potential of these tumors, high recurrence rates, and low treatment responses. Proteinases have been implicated in several aspects of tumor growth and metastasis in a broad range of tumors including HNSCC.Experimental Design: Comprehensive expression profiling of proteinases [matrix metalloproteinases (MMPs), A disintegrin and metalloproteinase (ADAMs), and ADAMs with thrombospondin motif (ADAMTSs)] and their inhibitors [tissue inhibitor of metalloproteinases (TIMPs)] was done using quantitative real-time reverse transcription-PCR analysis of a large cohort of tissue samples representing the tumor (n = 83), the invasive margin (n = 41), and the adjacent tissue (n = 41) from 83 HNSCC patients, along with normal tissue controls (n = 13), as well as cell lines established from tumors of 34 HNSCC patients.Results: The results show specifically elevated gene expression of several proteinases, including MMP1, MMP3, MMP10, and MMP13 within tumor tissue and peritumoral adjacent tissue. In addition, the results identify several novel HNSCC-associated proteinases, including ADAM8, ADAM9, ADAM17, ADAM28, ADAMTS1, ADAMTS8, and ADAMTS15. There were also significant differences in proteinase expression based on clinical parameters, i.e., tumor location, grade, and local invasion. MMP13 expression was significantly higher in large (>4 cm) locally invasive tumors (P < 0.05). MMP9 expression was significantly decreased in tumors with regional metastasis, whereas increased expression of ADAM8 was noted in the metastatic tumors (P < 0.001 for both).Conclusions: These findings suggest the HNSCC degradome as a valuable source of diagnostic, predictive, and prognostic molecular markers for these malignant tumors. Clin Cancer Res; 16(7); 2022-35. ©2010 AACR.Head and neck squamous cell carcinoma (HNSCC) can be defined as a malignant tumor derived from the squamous epithelial cells that line the upper aerodigestive tract, which extends from the surface of the lips to the cervical surface of the esophagus. HNSCC is the fifth most common cancer worldwide and, in 2002, was the cause of over 300,000 deaths worldwide (1). The overall survival rates of HNSCC patients (∼50%) have remained unaltered for over 20 years. The presence of lymph node metastases is the single most important predictive factor for HNSCC patients resulting in a 50% decrease in survival (2, 3).The degradome comprises both the complete set of proteases (also known as peptidases and proteinases) expressed by a cell, tissue, or organism at a given time, and the substrate repertoire of a given proteinase (4). There is a well-established association between matrix proteolysis and cancer invasion and metastasis, and the proteinases of every class have been linked to malignancy and invasion of tumor cells (5). However, through the study of transgenic models and the identification of further proteinases and proteinase s...

Serpin Peptidase Inhibitor Clade A Member 1 (SerpinA1) Is a Novel Biomarker for Progression of Cutaneous Squamous Cell Carcinoma

“…19 -21 SCC cells were cultured in Dulbecco's modified Eagle's medium supplemented with 6 mmol/L glutamine, nonessential amino acids, and 10% fetal calf serum. 20,21 HaCaT, a spontaneously immortalized, nontumorigenic human epidermal keratinocyte-derived cell line, 22 and the Ha-rastransformed tumorigenic HaCaT cell lines A5, II4, and RT3 23,24 (a gift from Dr. Norbert E. Fusenig, German Cancer Research Center, Heidelberg, Germany) were cultured in Dulbecco's modified Eagle's medium containing 10% fetal calf serum. G418 (200 g/mL) was added to the medium of the Ha-ras-transformed HaCaT cell lines.…”

“…Keratinocytes were cultured in keratinocyte basal medium 2, supplemented with SingleQuots (Cambrex Bioscience, Walkersville, MD), as previously described. 20,21 …”

“…cDNA was reverse transcribed from 1 g of RNA using reverse transcriptase M-MLV RNase H without reverse transcriptase and random hexamers (Promega, Helsinki, Finland). 20,21 …”

“…20,21 The levels of ␤-actin and glyceraldehyde-3-phosphate dehydrogenase mRNA (as housekeeping genes) were determined as described previously. 21 …”

Molecular discrimination of cutaneous squamous cell carcinoma from actinic keratosis and normal skin

“…7,8 Upregulation of MMP1 appears to a conserved in squamous cell carcinomas from different anatomic locations including the head and neck and oral cavity. [67][68][69][70] Matrix metallopeptidases appear integral to tumorigenesis and have been targeted by inhibitors; however, clinical trials of advanced lung, prostate, pancreas, brain, and GI tract malignancies have shown little efficacy to date. 64,71 More data may be needed before a successful clinical trial is completed.…”