sq-23377 and Head-and-Neck-Neoplasms

The present study aims to further explore the role of STK33 in hypopharyngeal squamous cell carcinoma (HSCC), with special attention given to the possible relationship between STK33 alteration and calcium.. An in vivo experiment and microarray analysis were performed to investigate the impact of STK33 knockdown (STK33-RNAi) on the biological behaviors and the gene profile alterations of a HSCC cell line (Fadu). Cell viability and morphological change of Fadu cells in response to Ionomycin were measured by MTT assay and acridine orange staining. The concentration of intracellular calcium ([Ca(2+)]i) was detected by laser scanning confocal microscope with fluo-3/AM. The mRNA and protein expressions of relevant genes were examined by real-time PCR and Western blot.. STK33-RNAi retarded the Fadu cell proliferation and the metastasis in nude mice and led to up- and down-regulation of the expressions of abundance of genes, especially, the downregulation of the CAPN1 gene. Ionomycin increased the [Ca(2+)]i and decreased the survival rates of Fadu cells in a time-dependent manner. Moreover, Ionomycin resulted in the elevation of CAPN1 mRNA expression in normal Fadu cells and, conversely, had almost no effect on CAPN1 expression in STK33-RNAi cells.. Findings from this work further validate that STK33 is a potential oncogene and plays an important role in tumorigenesis of HSCC via regulation of numerous genes. In addition, there exists the reciprocal influence between STK33 and [Ca(2+)]i in Fadu cells.

Topics: Animals; Calcium; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Gene Knockdown Techniques; Head and Neck Neoplasms; Heterografts; Humans; Hypopharyngeal Neoplasms; Ionomycin; Male; Mice; Mice, Nude; Protein Serine-Threonine Kinases; Random Allocation; RNA, Small Interfering; Squamous Cell Carcinoma of Head and Neck

Despite lots of research efforts, the pathology of head and neck cancer remains elusive. Accumulating evidence suggests that the innate and adaptive immunity plays an important role in HNSCC (Head and Neck Squamous Cell Carcinoma) development. Recently, a new T helper cell subset additional to the classical Th1 and Th2 cells was identified called Th17 cells, due to their secretion of IL-17. However, Th17 cells also produce additional proinflammatory cytokines and many other cytokines are involved in their differentiation and expansion. It was shown that Th17 cells play a prominent role in host defense but are also associated with the development of autoimmune diseases. The role of Th17 cells in cancer pathogenesis remains nebulous.. Th17 cells of peripheral blood, primary tumors and metastatic lymph nodes were FACS analyzed for their CD161 expression. Supernatants of the permanent HNSCC cell line BHY were used to induce Th17 cells by HNSCC tumor mileu.. Here we show that Th17 cells from patients with HNSCC downregulate the Th17 cell surface receptor CD161 in peripheral blood as well as in primary tumors and especially in metastatic lymph nodes.. We have showed for the first time alterations of Th17 cell phenotype in HNSCC patients.

Topics: Aged; Brefeldin A; Carcinoma, Squamous Cell; Culture Media, Conditioned; Head and Neck Neoplasms; Humans; Ionomycin; Leukocytes, Mononuclear; Lymph Nodes; Lymphocyte Activation; Middle Aged; Neoplasm Metastasis; NK Cell Lectin-Like Receptor Subfamily B; Tetradecanoylphorbol Acetate; Th17 Cells

A subset of circulating T cells (CD8(+)CD45RO(-)CD27(-)) with a naïve phenotype, but mediating effector function, is considered to play an important role in host antitumour defence. To investigate the attributes of these effector T cells in patients with squamous cell carcinoma (SCC) of the head and neck cancer, venous blood was obtained from 39 individuals with cancer and 45 normal controls (NC). Peripheral blood mononuclear cells were isolated, stained with labelled monoclonal antibodies specific for CD8, CD45RO, CD45RA, CD62L, CD27, TCR-zeta as well as isotype controls and examined by multicolour flow cytometry. Annexin V binding to CD8(+) T cells and PMA/ionomycin-induced IFN-gamma expression were also evaluated in patients and NC. The proportions of CD45RA(+)CD45RO(-) (naïve) and CD45RA(-)CD45RO(+) (memory) cells were found to be comparable within the CD8(+) T-cell subset. However, relative to NC, the frequency of effector CD8(+)CD45RO(-)CD27(-) cells was strikingly increased in all SCC patients regardless of the disease status (P=0.0003). The proportion of these cells was found to increase with age in both patients and NC. In NC, stimulated IFN-gamma expression was largely restricted to CD8(+)CD45RO(-)CD27(+) cells, while in patients CD8(+)CD45RO(-)CD27(-) expressed IFN-gamma after ex vivo stimulation. Expression of the TCR-associated zeta chain was decreased or absent in freshly isolated CD8(+)CD45RO(-)CD27(-) T cells in patients (P<0.0001). Annexin V was found to bind to a higher proportion of circulating CD8(+) T cells in patients than NC (P<0.006), and significantly more Annexin V(+) T cells were present in the effector (P<0.0059) than the naïve subset within the CD8(+)CD45RO(-) compartment. The data indicate that the expanded CD8(+)CD45RO(-)CD27(-) T cells, which contain precursors of IFN-gamma-producing T cells, are zeta-negative and sensitive to apoptosis in the circulation of patients with HNC.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Annexin A5; Apoptosis; Carcinogens; Carcinoma, Squamous Cell; Case-Control Studies; CD8-Positive T-Lymphocytes; Cohort Studies; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; Immunologic Memory; Interferon-gamma; Ionomycin; Ionophores; Leukocyte Common Antigens; Male; Middle Aged; Receptors, Antigen, T-Cell; T-Lymphocyte Subsets; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor Receptor Superfamily, Member 7

Fifty-one cervical nodes from 19 patients with advanced head and neck cancer were stimulated with phorbol dibutyrate and ionomycin (PDBu + Io) to determine the effect of such stimulation on the generation of cytotoxic T cells and whether this stimulation could bypass the need for autologous tumor stimulation. Lymphocytes stimulated with PDBu + Io demonstrated a sixfold greater in vitro expansion and significantly increased DNA synthesis. Whereas fresh lymphocytes displayed no cytotoxicity, stimulation with PDBu + Io and culture in interleukin-2 (IL-2) led to significant cytotoxicity equivalent to that of lymphocytes stimulated with autologous tumor and IL-2. T cells with the greatest cytotoxicity were generated from patients with nodal metastases. In patients with stage IV tumors, effector cells demonstrating greater lysis of natural killer-resistant targets (Daudi cells) were associated with higher rates of recurrence (50% versus 12%, respectively, p < 0.001). Stimulation with PDBu + Io augments growth and proliferation of lymphocytes from draining lymph nodes and preserves cytotoxicity without the need for autologous tumor. Excluding the need for antigenic stimulation by autologous tumor may prove useful in adoptive immunotherapy procedures.

Topics: Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; Cell Division; Cytotoxicity Tests, Immunologic; Head and Neck Neoplasms; Humans; Immunotherapy, Adoptive; Ionomycin; Lymph Nodes; Lymphatic Metastasis; Lymphocyte Activation; Phorbol 12,13-Dibutyrate; T-Lymphocytes, Cytotoxic; Tumor Cells, Cultured