Protein Expression in Tonsillar and Base of Tongue Cancer and in Relation to Human Papillomavirus (HPV) and Clinical Outcome

Abstract

Human papillomavirus (HPV) is a major etiological factor for tonsillar and the base of tongue cancer (TSCC/BOTSCC). HPV-positive and HPV-negative TSCC/BOTSCC present major differences in mutations, mRNA expression and clinical outcome. Earlier protein studies on TSCC/BOTSCC have mainly analyzed individual proteins. Here, the aim was to compare a larger set of cancer and immune related proteins in HPV-positive and HPV-negative TSCC/BOTSCC in relation to normal tissue, presence of HPV, and clinical outcome. Fresh frozen tissue from 42 HPV-positive and 17 HPV-negative TSCC/BOTSCC, and corresponding normal samples, were analyzed for expression of 167 proteins using two Olink multiplex immunoassays. Major differences in protein expression between TSCC/BOTSCC and normal tissue were identified, especially in chemo- and cytokines. Moreover, 34 proteins, mainly immunoregulatory proteins and chemokines, were differently expressed in HPV-positive vs HPV-negative TSCC/BOTSCC. Several proteins were potentially related to clinical outcome for HPV-positive or HPV-negative tumors. For HPV-positive tumors, these were mostly related to angiogenesis and hypoxia. Correlation with clinical outcome of one of these, VEGFA, was validated by immunohistochemistry. Differences in immune related proteins between HPV-positive and HPV-negative TSCC/BOTSCC reflect the stronger activity of the immune defense in the former. Angiogenesis related proteins might serve as potential targets for therapy in HPV-positive TSCC/BOTSCC.

Protein profiling of fine‐needle aspirates reveals subtype‐associated immune signatures and involvement of chemokines in breast cancer

“…Notably, among the top 11 proteins expressed at higher levels in the ‘ER High & Ki67 Low’ sample cluster, seven proteins were also expressed at elevated levels in HPV‐positive tonsillar and base of tongue cancer vs normal adjacent tissues (CA9, CXCL10, MMP12, CCL20, CXCL11, CXCL9, and CXCL8; Ramqvist et al ., ). This indicates striking similarities in immune activation between the diverse tumor types.…”

Immune related proteins and tumor infiltratingCD8+ lymphocytes in hypopharyngeal cancer in relation to human papillomavirus (HPV) and clinical outcome

“…Samples were prepared for protein analysis essentially as described in Ramqvist et al 27 Briefly, six cuts were made from each biopsy (1 × 5 μm, 4 × 20 μm, and 1 × 5 μm), frozen, and embedded in optimal cutting temperature compound. The first and last slides were used for evaluation of tumor content by an experienced pathologist and only tumors with ≥40% tumor cells were included in the protein evaluation and 28/33 samples included in the analysis had ≥70% tumor cells.…”

“…All normal samples were checked to be free from tumor tissue. The four 20 μm cuts from each sample were pooled, dissolved in RIPA buffer, and frozen at −70°C until analysis on the Proseek panel as described in Ramqvist et al 27 …”

“…Recently, we investigated protein expression in OPSCC in relation to normal tissue, HPV status, and clinical outcome, utilizing Proseek multiplex panels (Olink Bioscience, Uppsala, Sweden) based on proximity extension assay (PEA) technology, and found an increased expression of many immune related proteins in HPV‐positive vs HPV‐negative tumors 27 . In the present study, we aimed to, with the same method, investigate the expression of immune‐related proteins in HPSCC in relation to normal tissue, and tumor HPV‐status.…”

Effects of flavonoids on tongue squamous cell carcinoma

“…Initially, patients receive prolonged high‐dose radiation therapy for 6–7 weeks, and stage III or IV patients also receive chemotherapy and EGFR blockers (Lindquist et al, ; Kraaijenga et al, ), which cause a wide range of side effects, such as pain, swallowing and ingestion difficulties, nausea, infections, mucositis, and fatigue. Patients with involved lymph nodes undergo cervical lymph node removal, resulting in increased neck rigidity, limited shoulder mobility, xerostomia, altered sense of taste, hearing problems, and on occasion osteonecrosis requiring reconstructive surgery (Dai et al, ; Loorents et al, ; Ramqvist et al, ). Cetuximab (e.g., Erbitux®) can be used, but it can increase pain and is unaffordable for many patients (Dai et al, ).…”