bromochloroacetic-acid and Squamous-Cell-Carcinoma-of-Head-and-Neck

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Proliferation; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Melanoma; Nose Neoplasms; S100 Proteins; Skin Neoplasms; Squamous Cell Carcinoma of Head and Neck

In head and neck squamous cell carcinoma (HNSCC), salvage neck dissection (ND) is required after primary chemoradiation in case of residual nodal disease. Upon histopathological examination, viability of tumor cells is assessed but little is known about other prognostic histopathological features. In particular, the presence of swirled keratin debris and its prognostic value is controversial. The aim of this study is to examine histopathological parameters in ND specimens and correlate them with patient outcome to determine the relevant parameters for histopathological reporting.. Salvage ND specimen from a cohort of n = 75 HNSCC (oropharynx, larynx, hypopharynx) patients with prior (chemo) radiation were evaluated on H&E stains for the following parameters: viable tumor cells, necrosis, swirled keratin debris, foamy histiocytes, bleeding residues, fibrosis, elastosis, pyknotic cells, calcification, cholesterol crystals, multinucleated giant cells, perineural, and vascular invasion. Histological features were correlated with survival outcomes.. Only the presence / amount (area) of viable tumor cells correlated with a worse clinical outcome (local and regional recurrence-free survival, (LRRFS), distant metastasis-free survival, disease-specific survival, and overall survival, p < 0.05) in both the univariable and multivariable analyses.. We could confirm the presence of viable tumor cells as a relevant negative prognostic factor after (chemo) radiation. The amount (area) of viable tumor cells further substratified patients with worse LRRFS. None of the other parameters correlated with a distinctive worse outcome. Importantly, the presence of (swirled) keratin debris alone should not be considered viable tumor cells (ypN0).

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratins; Lymphatic Metastasis; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Retrospective Studies; Squamous Cell Carcinoma of Head and Neck

Oral squamous cell carcinoma (OSCC) is one of the most common malignancies affecting the head-and-neck region, regional lymph nodes being an important prognostication factor dictating the survival rate. Despite an array of modalities used, clinically, radiographically and routine histopathologically, the detection of micro-metastasis (2-3 mm tumour cell deposits) in the lymph nodes often escapes identification. The presence of few of these tumour epithelial cells in the lymph nodes drastically increases mortality and alters treatment plan. Hence, the identification of these cells is of major prognostic significance for a patient. Thus, the present study was aimed to evaluate and detect the efficacy of the immunohistochemical (IHC) marker [cytokeratin (CK) AE1/AE3] over routine Hematoxylin & eosin (H & E) staining in detecting micro-metastasis in the lymph nodes of OSCC cases.. The IHC marker CK cocktail (AE1/AE3) did not demonstrate any positive reactivity for the target antigen in all the 100 H & E stained lymph node sections evaluated in the present study.. This study was undertaken to check the efficacy of IHC (CK cocktail AE1/AE3) in the detection of micro-metastasis in lymph nodes that are found to be negative in routine H&E stained sections. The findings of this study suggest that the IHC marker AE1/AE3 did not prove to be useful to detect micro-metastasis in this study population.

Topics: Carcinoma, Squamous Cell; Eosine Yellowish-(YS); Head and Neck Neoplasms; Hematoxylin; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

We investigated whether in human head and neck squamous cell carcinoma (HNSCC) high levels of expression of stress keratin 17 (K17) are associated with poor survival and resistance to immunotherapy.. We investigated the role of K17 in regulating both the tumor microenvironment and immune responsiveness of HNSCC using a syngeneic mouse HNSCC model, MOC2. MOC2 gives rise to immunologically cold tumors that are resistant to immune-checkpoint blockade (ICB). We engineered multiple, independent K17 knockout (KO) MOC2 cell lines and monitored their growth and response to ICB. We also measured K17 expression in human HNSCC of patients undergoing ICB.. MOC2 tumors were found to express K17 at high levels. When knocked out for K17 (K17KO MOC2), these cells formed tumors that grew slowly or spontaneously regressed and had a high CD8+ T-cell infiltrate in immunocompetent syngeneic C57BL/6 mice compared with parental MOC2 tumors. This phenotype was reversed when we depleted mice for T cells. Whereas parental MOC2 tumors were resistant to ICB treatment, K17KO MOC2 tumors that did not spontaneously regress were eliminated upon ICB treatment. In a cohort of patients with HNSCC receiving pembrolizumab, high K17 expression correlated with poor response. Single-cell RNA-sequencing analysis revealed broad differences in the immune landscape of K17KO MOC2 tumors compared with parental MOC2 tumors, including differences in multiple lymphoid and myeloid cell types.. We demonstrate that K17 expression in HNSCC contributes to immune evasion and resistance to ICB treatment by broadly altering immune landscapes of tumors.

Topics: Animals; Head and Neck Neoplasms; Humans; Immune Checkpoint Inhibitors; Immune Evasion; Keratin-17; Keratins; Mice; Mice, Inbred C57BL; Squamous Cell Carcinoma of Head and Neck; Tumor Microenvironment

Extracellular vesicles (EVs) have recently come into the spotlight as potential cancer biomarkers. Isolation of pure EVs is complex, so wider use requires reliable and time‑efficient isolation methods. In the present study, galectin‑based magnetic glycan recognition particles, EXÖBead

Topics: B7-H1 Antigen; Biomarkers, Tumor; Epithelial Cell Adhesion Molecule; Extracellular Vesicles; Galectins; Head and Neck Neoplasms; Humans; Keratins; Leukocytes, Mononuclear; Lipoproteins; Polyethylene Glycols; Polysaccharides; Squamous Cell Carcinoma of Head and Neck

Fanconi anemia (FA) patients frequently develop oral squamous cell carcinoma (OSCC). This cancer in FA patients is diagnosed within the first 3-4 decades of life, very often preceded by lesions that suffer a malignant transformation. In addition, they respond poorly to current treatments due to toxicity or multiple recurrences. Translational research on new chemopreventive agents and therapeutic strategies has been unsuccessful partly due to scarcity of disease models or failure to fully reproduce the disease. Here we report that Fanca gene knockout mice (Fanca

Topics: Animals; Carcinoma, Squamous Cell; Disease Models, Animal; Fanconi Anemia; Head and Neck Neoplasms; Keratins; Mice; Mice, Knockout; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

The TSA Opal multiplex immunohistochemistry (mIHC) protocol (PerkinElmer) has been used to characterize immune infiltration in human cancers. This technique allows multiple biomarkers to be simultaneously stained in a single tissue section, which helps to elucidate the spatial relationship among individual cell types. We developed and optimized two improved mIHC protocols for a 7-color panel containing 6 biomarkers (CD3, CD8, CD163, PD-L1, FoxP3, and cytokeratin (CK)) and DAPI. The only difference between these two protocols was the staining sequence of those 6 biomarkers as the first sequence is PD-L1/CD163/CD8/CK/CD3/FoxP3/DAPI and the second sequence is FoxP3/CD163/CD8/CK/CD3/PD-L1/DAPI. By comparing PD-L1/FoxP3 staining in mIHC and singleplex PD-L1/FoxP3 staining on the adjacent slide, we demonstrated that the staining sequence does not affect the staining intensity of individual biomarkers as long as a proper antigen retrieval method was used. Our study suggests that use of an antigen retrieval buffer with higher pH value (such as Tris-EDTA pH9.0) than that of the stripping buffers (such as citrate buffer pH6.0) is helpful when using this advanced mIHC method to develop panels with multiple biomarkers. Otherwise, individual biomarkers may exhibit different intensities when the staining sequence is changed. By using this protocol, we characterized immune infiltration and PD-L1 expression in head and neck squamous cell carcinoma (HNSCC), breast cancer (BCa), and non-small cell lung cancer (NSCLC) specimens. We observed a statistically significant increase in CD3+ cell populations within the stroma of NSCLC as compared to BCa and increased PD-L1+ tumor cells in HNSCC as opposed to BCa.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; B7-H1 Antigen; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Non-Small-Cell Lung; CD3 Complex; CD8 Antigens; Female; Forkhead Transcription Factors; Head and Neck Neoplasms; Humans; Immunohistochemistry; Indoles; Keratins; Lung Neoplasms; Lymphocytes, Tumor-Infiltrating; Receptors, Cell Surface; Squamous Cell Carcinoma of Head and Neck

Topics: Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Esophagoscopy; Humans; Keratins; Squamous Cell Carcinoma of Head and Neck

The role of Cytokeratin fraction 21-1 (CYFRA 21-1) as a tumour marker for head and neck cancer is still a matter of research. The aim of the present study was to evaluate the clinical impact of CYFRA 21-1 for patients with oropharyngeal squamous cell carcinoma (OSCC).. Data of 180 patients with an initial diagnosis of OSCC of any stage between 2003 and 2017 were retrospectively analysed regarding the association between pretherapeutic CYFRA 21-1 levels, clinical characteristics, overall and disease-free survival. Additionally, the potential of CYFRA 21-1 for the detection of recurrent disease in the follow-up was evaluated. The cut-off value was set at 3.3 ng/ml. The median follow-up time was 2.85 years.. A significant correlation of the CYFRA 21-1 concentration at the time of diagnosis and the N-stage was detected (p = 0.01). Patients with CYFRA 21-1 levels > 3.3 ng/ml at first diagnosis showed a significantly shorter overall survival. In the case of disease-progression, a significant increase of CYFRA 21-1 value was found compared to post-therapeutic CYFRA 21-1 levels (9.1 ng/ml versus 5.1 ng/ml; p < 0.01). CYFRA 21-1 level after treatment showed only a low sensitivity of 32% and a specificity of 78% for tumour recurrence.. CYFRA 21-1 correlates with the tumour stage and, therefore, the survival of OSCC patients. Posttreatment CYFRA21-1 seems not to be a suitable predictor of tumour recurrence in the further course of the disease. However, a sudden increase of CYFRA 21-1 during follow-up may indicate a tumour recurrence in the individual patient.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Recurrence, Local; Retrospective Studies; Sensitivity and Specificity; Squamous Cell Carcinoma of Head and Neck

Tumor budding (TB) is a promising prognostic marker in many cancers including oral squamous cell carcinoma. The evaluation of TB in preoperative diagnostic biopsies has been proven be possible; therefore, the association of TB with other morphological features can represent an important aid in the previous treatment decision. This study aims to evaluate TB in oral squamous cell carcinoma (OSCC) biopsies, assessing its association with other morphological characteristics of the sample. A total of 56 cases of OSCC were investigated. In hematoxylin and eosin-stained slides, morphological features including histopathological grading and mode of invasion were evaluated in the deep invasive front. Moreover, immunohistochemistry was performed with anti-multi-cytokeratin antibody helping in the identification of TB, which was graded as low-intensity or no TB and high-intensity TB. Descriptive and bivariate analyses were performed, and the level of significance was set at 5%. The tongue was the most-affected site with 29 (51.7 %) tumors. The predominant mode of invasion (27-48.2 %) was by groups of neoplastic cells without clear boundaries. Of the cases investigated, 37 (66.1 %) were high-intensity TB, which was associated with the mode of invasion of the tumors (p < 0.05). All cases with the worst mode of invasion showed high-intensity TB. Preliminary results showed the potential of morphological features, such as TB and mode of invasion, evaluated in diagnostic specimens of OSCC, aiding in the treatment decision to select patients who could benefit from more-aggressive treatments.

Topics: Adult; Biopsy; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

Occult neck metastasis is an important prognostic factor in patients with tongue squamous cell carcinoma (TSCC) who are deemed clinically negative for neck metastasis. The purpose of this study was to identify predictive factors for occult neck metastasis arising from TSCC and to determine patient prognosis. Ninety-seven patients with cT2N0 TSCC who underwent surgical resection of their primary lesion as initial therapy were enrolled in this retrospective study. Cutoff values for depth of invasion (≥3.3 mm) and the tumor budding score (≥4) were determined using receiver operator characteristic analyses. Univariate and multivariate analyses revealed that a tumor budding score ≥4 is a significant independent predictive factor for the occurrence of occult neck metastasis, which in turn is a significant independent prognostic factor. When evaluating tumor budding, we demonstrated greater interobserver and intraobserver agreement when using immunohistochemical staining for cytokeratin AE1/AE3 than with hematoxylin and eosin staining (HE). We conclude that the evaluation of tumor budding is effective for identifying populations at high risk of occult neck metastasis, which will enable the planning of appropriate therapeutic strategies for patients with cT2N0 TSCC. Furthermore, cytokeratin staining is recommended over HE staining for simpler and more accurate evaluation of tumor budding.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cell Movement; Female; Glossectomy; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Observer Variation; Predictive Value of Tests; Reproducibility of Results; Retrospective Studies; Squamous Cell Carcinoma of Head and Neck; Staining and Labeling; Tongue Neoplasms; Treatment Outcome

High-risk human papilloma virus (HPV) testing should be performed on all patients with newly diagnosed oropharyngeal HPV-associated squamous cell carcinoma (OPHPVSCC), and p16 immunostaining can be used as a surrogate marker. Although in surgical pathology specimens p16 staining in > 70% of the tumor cells is considered a positive result, the interpretation in fine-needle aspiration (FNA) specimens has remained controversial.. FNA of neck lymph nodes and corresponding surgical specimens from 42 patients with OPHPVSCC were reviewed.. In FNA specimens, 38 cases (90.5%) had viable tumor cells, 32 (76.2%) had keratin debris, and 36 (85.7%) had degenerated keratinized tumor cells. Twenty-seven of 27 (100%) had positive p16 staining in > 70% of viable tumor cells, while the degenerated tumor cells were negative. Twenty of 24 (83.3%) primary OPHPVSCC exhibited focal degenerated keratinized tumor cells and/or keratin debris.. This study showed that the majority of the OPHPVSCC metastases in lymph nodes had degenerated keratinized tumor cells and keratin debris. Many primary OPHPVSCC also demonstrated focal keratinization and/or degeneration. The degenerated tumor cells showed no immunoreactivity to p16. The same 70% cutoff used in histologic specimens should be applied in cytologic specimens, but only the viable tumor cells should be counted.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy, Fine-Needle; Cell Survival; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Male; Middle Aged; Oropharyngeal Neoplasms; Papillomavirus Infections; Predictive Value of Tests; Reproducibility of Results; Squamous Cell Carcinoma of Head and Neck

Squamous cell carcinoma (SSC) of the head and neck is the sixth most common cancer and is rarely diagnosed in early stages. The transcription factor Krϋppel-like factor 4 (Klf4) suppresses cell proliferation and promotes differentiation. Inducible mice carrying an oral-specific ablation of Klf4 (K14-CreER(tam) /Klf4(flox/flox) ) develop mild dysplastic lesions and abnormal differentiation in the tongue. Aiming to analyze whether Klf4 cooperate in oral chemical carcinogenesis,we applied 4-nitroquinoline 1-oxide (4NQO), a tobacco surrogate, to this conditional Klf4 knockout mice.. K14-CreER(tam) /Klf4(flox/flox) and control mice were treated with 4NQO for 16 weeks and monitored until week 30. Histopathological samples were used for diagnostic purposes and immunofluorescence detection of epithelial differentiation markers.. 4NQO-treated K14-CreER(tam) /Klf4(flox/flox) mice (Klf4KO 4NQO) showed a significant weight loss and developed more severe dysplastic lesions than control mice with 4NQO (P < 0.005). The Klf4KO 4NQO showed a tendency to higher incidence of oral SCC and a marked keratinization pattern in dysplasias, in situ carcinomas and SCC. Also, tongues derived from Klf4KO 4NQO mice exhibited reduced terminal differentiation as judged by cytokeratin 1 staining when compared with 4NQO-treated controls.. Klf4 ablation results in more severe dysplastic lesions in oral mucosa, with a tendency to higher incidence of SCC, after chemical carcinogenesis. We show here, in a context similar to the human carcinogenesis, that absence of Klf4 accelerates carcinogenesis and correlates with the absence of cytokeratin 1 expression. These results suggest a potential role for KLF4 as a tumor suppressor gene for the tongue epithelium.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Carcinogens; Carcinoma, Squamous Cell; Cell Differentiation; Disease Models, Animal; Gene Expression; Head and Neck Neoplasms; Keratins; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Mice; Mice, Inbred C57BL; Mice, Knockout; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Squamous Cell Carcinoma of Head and Neck; Tongue Neoplasms

The invasion of cancer cells into the surrounding normal tissue is one of the defining features of cancer. While the phenomena of tumour budding, epithelial-mesenchymal transition and the presence of myofibroblasts have independently been shown to be related to a poor prognosis of oral carcinomas, their relationship has not been examined in detail.. Paraffin-embedded tissues from 28 patients with oral squamous cell carcinomas were stained with antibodies to cytokeratin, α-SMA, vimentin, E-cadherin, N-cadherin and Twist and evaluated for their expression in relation to invasive cancer cells and the surrounding tumour stroma.. A direct, histological relationship between invading, budding tumour cells and myofibroblasts was occasionally seen but was not a general feature. Most of the budding tumour cells at the invasive front had a decreased expression of E-cadherin, but we did not find that this was associated with a consistent or clear increase in either N-cadherin or vimentin. We therefore suggest that the budding of tumour cells is not dependent upon either myofibroblasts or a complete epithelial-mesenchymal transition and that these phenomena most likely represent separate processes in tumour progression.

Topics: Actins; Animals; Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Goats; Head and Neck Neoplasms; Humans; Keratins; Mice; Mouth Neoplasms; Muscle, Smooth; Myofibroblasts; Paraffin Embedding; Prognosis; Rabbits; Single-Cell Analysis; Squamous Cell Carcinoma of Head and Neck; Twist-Related Protein 1; Vimentin

The objective of this study was to evaluate the potential detection of circulating tumor cells (CTCs) using the CellSearch (CS) Assay™ in patients with locally advanced head and neck squamous cell carcinoma (HNSCC) and then to identify the clinical factors predictive of the presence of CTCs. The presence and number of CTCs were determined using the CS system before treatment, and in 10 healthy individuals (control group). The CS system was able to successfully identify the presence of CTCs in 8 of 49 patients (16 %) before therapy. No CTC was found in the control group. CTCs were detected before therapy in 1 of 19 patients (5 %) with N0 tumor and in 7 of 30 patients (23 %) with N1-2c tumor (p = 0.12; Fisher's exact test). CTCs were identified in a relatively low proportion of patients with locally advanced HNSCC.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Cell Adhesion Molecules; Cell Count; Epithelial Cell Adhesion Molecule; Female; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Neoplastic Cells, Circulating; Oropharyngeal Neoplasms; Prognosis; Squamous Cell Carcinoma of Head and Neck

The objective of this study is to evaluate the incidence of occult nodal micrometastases in N0 laryngeal squamous cell carcinoma using cytokeratin immunohistochemical analysis (CKIHA) and its influence on the overall occult metastatic rate. This is a prospective cohort study. A total number of 30 patients with N0 stage laryngeal cancer underwent 46 selective neck dissections for elective treatment of the neck. Nodes found to be negative using routine histopathological examination were evaluated for the presence of micrometastasis using CKIHA. The occult micrometastasis rate for all cases was 26.7 % which significantly increased the overall occult metastasis rate to 50 % (P = 0.014). The micrometastasis rate was 30.8, 25 and 20 % for glottic, supraglottic and transglottic tumors, respectively, which increased the overall occult metastasis rate to 53.8, 50 and 40 % but without statistical impact. The micrometastasis rate was 35.7 and 23.1 % in T3 and T4 tumors, respectively, and this increased the overall occult metastasis rate to 50 and 61.5 % with statistical influence in T3 tumors (P = 0.046). Micrometastasis upstaged the neck in 7 (23.3 %) patients with statistical impact on the PN stage (P = 0.007). The overall occult nodal metastasis rate in N0 laryngeal cancer is underestimated. Nodal micrometastasis may be missed during routine histopathological examination and can be detected using CKIHA.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cohort Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Neck Dissection; Neoplasm Micrometastasis; Neoplasm Staging; Prospective Studies; Sensitivity and Specificity; Squamous Cell Carcinoma of Head and Neck

The migratory ability of tumor cells requires cytoskeletal rearrangement processes. Epidermal growth factor receptor (EGFR)-signaling tightly correlates with tumor progression in head and neck squamous cell carcinomas (HNSCCs), and has previously been implicated in the regulation of cytokeratin (CK) expression. In this study, HNSCC cell lines were treated with EGF, and CK expression levels were monitored by Western blot analysis. Changes in cellular morphology were documented by fluorescence- and atomic force microscopy. Some of the cell lines demonstrated an EGF-dependent modulation of CK expression levels. Interestingly, regression of some CK subtypes or initial up-regulation followed by downregulation at higher EGF-levels could also be observed in the tested cell lines. Overall, the influence of EGF on CK expression levels appeared variable and cell-type-dependent. Real-time cellular analysis of EGF-treated and -untreated HNSCC cell lines demonstrated a rise over time in cellular impedance. In three of the EGF-treated HNSCC cell lines, this rise was markedly higher than in untreated controls, whereas in one of the cell lines the gain of cellular impedance was paradoxically reduced after EGF treatment, which was found to correlate with changes in cellular morphology rather than with relevant changes in cellular viability or proliferation. After treating HNSCC cells with EGF, CK filaments frequently appeared diffusely distributed throughout the cytoplasm, and in some cases were found in a perinuclear localization, the latter being reminiscent to observations by other groups. In summary, the data points to a possible role of EGFR in modulating HNSCC cell morphology.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Shape; Cell Survival; Epidermal Growth Factor; Head and Neck Neoplasms; Humans; Keratinocytes; Keratins; Microscopy, Atomic Force; Phenotype; Plakophilins; Squamous Cell Carcinoma of Head and Neck

To determine the feasibility of viable storage of head and neck squamous cell carcinoma (HNSCC) for regrowth of cells in culture.. Laboratory-based translational study.. Methods for intermediate-term frozen storage of viable HNSCC were explored using small pieces of primary tumor and dissociated HNSCC cells after short-term culture. Viable cells after freezing were confirmed by adherence to tissue culture plates, cell morphology, and increased cell or colony density. Two cultures were immunostained for cytokeratin to confirm epithelial origin of viable cultured cells after freezing.. Six primary HNSCCs (two oral cavity, three larynx, one oropharynx) and two HNSCCs that had been passaged through a xenograft (two oral cavity) were dissociated to single cells and grown in short-term cell culture for 0 to 12 passages. After short-term culture, cells were frozen for up to 8 months, thawed, and replated. Frozen cells derived from all tumors (six primary and two xenografts) were successfully replated with cultures lasting >7 days with seven of eight tumors presenting increased colony or cell density over 1 week of growth after freezing. In total, 15 of 15 tested samples derived from six primary and two xenografted HNSCCs were viable after freezing.. In the current study, we show that biopreservation of primary or xenografted HNSCC using short-term cell culture is feasible. Initial short-term cell culture was required for successful storage and viability of frozen cells. These proof-of-principle studies, if more widely implemented, could improve preclinical testing of new therapies for HNSCC.

Topics: Carcinoma, Squamous Cell; Cryopreservation; Feasibility Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Mouth Neoplasms; Oropharyngeal Neoplasms; Squamous Cell Carcinoma of Head and Neck; Tissue Banks; Transplantation, Heterologous; Tumor Cells, Cultured

A high lactate content in malignant head and neck cancer (Head and neck squamous cell carcinomas, HNSCC) is associated with a higher risk of metastatic spread and lower overall patient survival. However, until present, the underlying mechanisms are not clearly understood. Here, a systematic comparison of glucose metabolism in HNSCC and homologous normal tissue is presented for the first time.. The concentrations of glucose, lactate and ATP were measured in cryobiopsies of 29 human HNSCC and of 9 normal mucosa using bioluminescence imaging. The protein expression of lactate dehydrogenase (LDH) was analyzed by Western blotting.. Tumors own a higher content of lactate and LDH in comparison with normal tissues. However, within the tumor group, the grade of LDH expression shows substantially strong variation and overlap with normal values. Furthermore, LDH expression was not correlated with tumor lactate content. Investigating a small subpopulation, patients with a short-term survival had significantly higher tumor lactate levels compared to patients with long-term survival.. The data provide clear evidence of an enhanced glycolysis in tumors compared to normal tissue. This may partially but not completely attributable to an elevated expression of LDH. High tumor lactate levels may be predictive for restricted patient survival. In conclusion, lactate measurements, for example non-invasively with MRT, should be advanced for use in clinical routine as a supportive tool for tumor diagnosis and prognosis.

Topics: Adenosine Triphosphate; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Blotting, Western; Carcinoma; Carcinoma, Squamous Cell; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gingiva; Glucose; Glycolysis; Head and Neck Neoplasms; Humans; Keratins; L-Lactate Dehydrogenase; Lactic Acid; Luminescence; Male; Middle Aged; Neoplasms, Squamous Cell; Predictive Value of Tests; Prognosis; Squamous Cell Carcinoma of Head and Neck