CALCB rs3829222 T/T Genotype and Low Expression of CALCB Are High-Risk Factors for Adenoid Cystic Carcinoma of Salivary Gland.

OBJECTIVES: To investigate the relationship between polymorphisms of calcitonin-related peptide gene II (beta-calcitonin gene-related peptide (ßCGRP), CALCB) and serum CGRP levels in salivary adenoid cystic carcinoma. MATERIALS AND METHODS: Using the polymerase chain reaction (PCR) technique, the full-length amplification and genotype analysis of CALCB genes were performed in 39 patients with adenoid cystic carcinoma of salivary gland and 158 normal controls. The gene frequencies of major genotype of CALCB in adenoid cystic carcinoma of salivary gland and normal control group were analyzed. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate serum calcitonin gene-related peptide (CGRP) and its concentration of alpha and beta subtypes. RESULTS: Univariate logistic regression analysis showed that the CALCB rs2839222 T/T genotype was closely related to the occurrence of salivary adenoid cystic carcinoma, with a correlation coefficient of 3.89. CONCLUSIONS: The serum CGRP concentration in the salivary adenoid cystic carcinoma group was 1.56 times that of the normal control group. The αCGRP subtype was significant, which was 3.02 times that of the normal control. The polymorphism of ßCGRP gene is associated with genetic susceptibility to salivary adenoid cystic carcinoma, and serum CGRP and ßCGRP can be used as novel markers of salivary adenoid cystic carcinoma.

Treatment-induced changes of lymphocyte subsets in patients with adenoid cystic carcinoma of the head and neck.

PURPOSE: Adenoid cystic carcinoma (ACC) of the head and neck is a rare and highly malignant tumor, characterized by perineural growth and early distant metastases. The composition of immune cells in the peripheral blood and the tumor microenvironment is critical to tumor growth and control. However, little is known about the frequency and function of the relevant immune cell subsets in this entity. METHODS: In ACC patients (n = 11) and matched healthy donors (n = 11), the frequency of peripheral blood T and B cells was measured by flow cytometry at different treatment stages of disease (24 samples). Cells were further characterized by their expression of CCR7, PD-1, CD39 and CD73. Tumor-infiltrating lymphocytes (TIL) were analyzed by immunohistochemistry for ten patients and for three patients by flow cytometry. RESULTS: CD4+ T cells had significantly lower frequency after radiotherapy (RT). All other cell frequencies, including Treg, were stable through course of the disease. In B cells, CD73 was reduced after RT. CCR7 expression on T and B cells in patients with relapse/metastases (R/M) differed significantly from patients with active disease. PD-1 remained stable. Treg were more present in TIL compared to peripheral blood. CONCLUSION: Composition of lymphocyte subgroups behaves similar to squamous cell carcinoma in the head and neck, except for Treg, which remained stable. Nevertheless, the CD4+/Treg ratio was lower after RT, which could stand for an immunosuppressive effect in these patients. Therefore, it could be beneficial treating ACC with combined RT and immunomodulatory drugs.

Investigation of IL-33 serum levels in patients with benign and malignant salivary gland tumors.

BACKGROUND: Interleukin-33 (IL-33) has been recently discovered as an influential factor in the process of tumor immunity, and is presented in cancer pathogenesis. OBJECTIVE: This study aimed to determine the serum levels of IL-33 in patients with benign and malignant Salivary gland tumors (SGTs). METHODS: This descriptive cross-sectional study was performed on 47 samples of malignant SGTs including 18 mucoepidermoid carcinoma (MEC), 8 adenoid cystic carcinoma (ADCC), 21 malignant mixed tumor (MMT), and 14 benign pleomorphic adenoma (PA). A control group was considered consisting of 28 healthy subjects. The serum level of IL-33 was measured by using sandwich ELISA method. The data were statistically analyzed through Kruskal-Wallis and Mann-Whitney tests. RESULTS: The median concentration of IL-33 was 6.91 in malignant, 5.14 in benign, and 5.01 in healthy cases, with a statistically significant difference (P= 0.001). The median serum levels of IL-33 increased significantly in ADCC (7.15), MEC (7.03), and MMT (6.91) compared with the control group (5.01) (P< 0.05). The mean rank of MEC was significantly higher than PA (P= 0.01). IL-33 concentration was positively and significantly correlated with the tumor stage (P= 0.02) and tumor size (P= 0.03). CONCLUSIONS: IL-33 could be suggested as a novel biomarker to distinguish different types of SGTs.

Expression and clinical significance of MAGE and NY-ESO-1 cancer-testis antigens in adenoid cystic carcinoma of the head and neck.

BACKGROUND: Adenoid cystic carcinoma (ACC) of the head and neck is a rare but highly malignant tumor. Cancer-testis antigens (CTAs) represent an immunogenic family of cancer-specific proteins and thus represent an attractive target for immunotherapy. METHODS: Eighty-four cases of ACC were identified, the CTAs pan-Melanoma antigen (pan-MAGE; M3H67) and New York esophageal squamous cell carcinoma (NY-ESO-1; E978) were detected immunohistochemically (IHC) and correlated with clinical data. RESULTS: Expression of NY-ESO-1 was found in 48 of 84 patients (57.1%) and of pan-MAGE in 28 of 84 patients (31.2%). Median overall survival (OS) in NY-ESO-1 positive versus negative patients was 130.8 and 282.0 months (p = .223), respectively. OS in pan-MAGE positive versus negative patients was 105.3 and 190.5 months, respectively (p = .096). Patients expressing both NY-ESO-1 and pan-MAGE simultaneously had significantly reduced OS with a median of 90.5 months compared with 282.0 months in negative patients (p = .047). CONCLUSION: A significant fraction of patients with ACC show expression of the CTAs NY-ESO-1 and/or pan-MAGE with promising immunotherapeutic implications. © 2016 Wiley Periodicals, Inc. Head Neck 38: 1008-1016, 2016.

Estudo imunoistoquímico de componentes da Via Sonic Hedgehog, Stat3 e Mcm3 em tumores de glândula salivar / Immunohistochemical study components of Via Sonic Hedgehog, Stat3 and MCM3 in salivary gland tumors

O adenoma pleomórfico (AP), o carcinoma mucoepidermóide (CME) e o carcinoma adenóide cístico (CAC) representam tumores frequentes em glândula salivar. A via de sinalização Sonic Hedgehog (Hh) e o Transdutor de sinal e ativador da transcrição 3 (STAT3) desempenham funções importantes na proliferação celular, favorecendo o desenvolvimento tumoral e a proteína MCM3 tem sido considerada uma nova classe de marcadores de proliferação celular. Portanto, o presente trabalho propõe-se a estudar componentes da via Hh, bem como o STAT3 e o MCM3 em neoplasias de glândula salivar, na tentativa de adicionar informações sobre as características biológicas dessas neoplasias. Foram utilizados 9 casos de AP, 17 casos de CAC e 20 casos de CME e, por meio da técnica imunoistoquímica, realizou-se a detecção das seguintes proteínas: SHH, GLI1, SUFU, HHIP, STAT3 e MCM3. No AP, observou-se alta expressão citoplasmática de SHH e SUFU, e baixa expressão de STAT3 e MCM3. No CAC, observou-se alta expressão de GLI1, HHIP e STAT3 e baixa expressão de SHH, SUFU e MCM3. No CME, observou-se alta expressão de SHH, GLI1, SUFU e HHIP e baixa expressão de STAT3 e MCM3. Quando comparado entre os tipos tumorais, observou-se diferença estatisticamente significante para expressão de SHH (p=0.0064), STAT3 (p=0.0003) e MCM3 (p=0.0257)...

The pleomorphic adenoma (PA), mucoepidermoid carcinoma (MEC) and the adenoid cystic carcinoma (ACC) are common tumors arising from salivary glands. The Sonic Hedgehog signaling pathway (Hh) and signal transducer and activator of transcription 3 (STAT3) play important roles in cell proliferation, favoring tumor growth. The MCM3 protein has been considered as a novel class of cell proliferation markers. The aim of this investigation was to study components of the Hh pathway, as well as STAT3 and MCM3 in salivary gland neoplasms in an attempt to add information about the biological characteristics of these neoplasms. We used 9 cases of PA, 17 cases of ACC and 20 cases of MEC. Using immunohistochemistry, were investigated: SHH, GLI1, Sufu, HHIP, STAT3 and MCM3. In PA, there was high expression of cytoplasmic SHH and Sufu, and low expression of STAT3 and MCM3. In the ACC, there was high expression of GLI1, HHIP and STAT3 and low expression of SHH, SUFU and MCM3. In the MEC, we observed high expression of SHH, GLI1, SUFU and HHIP and low expression of STAT3 and MCM3. There was a statistically significant difference between SHH (p=0.0064), STAT3 (p=0.0003) and MCM3 (p=0.0257) when all tumors were compared...

Study on anti-metastasis of heparin derivatives as ligand antagonist of p-selectin.

Heparin has a potential value as therapeutic agents that block P-selectin-mediated cell adhesion and prevent tumor metastasis. However, the strong anticoagulant potency limits its applicability for the anti-metastasis activity. Carboxyl and sulfate groups of heparin are closely related to its anticoagulant activity, so seven kinds of heparin derivatives related to carboxyl and sulfate groups were prepared, and their effects on anti-metastasis as ligand antagonist of p-selectin were studied. The results showed that heparin, carboxyl reduction heparin, 2-O-desulfated heparin and N-desulfated- N-acetylated heparin could inhibit the adhesion of tumor cells to endothelial cells and platelets effectively as ligand antagonist of P-selectin.

Serum soluble CD44v6 levels in patients with oral and maxillofacial malignancy.

OBJECTIVE: To determine the levels of serum sCD44v6 in patients with oral cancer and evaluate the value of serum sCD44v6 in adjuvant diagnosis, staging and monitoring treatment response in these patients. MATERIALS AND METHODS: A total of 112 hospitalized patients with oral and maxillofacial malignancy and 28 healthy individuals were examined for serum sCD44v6 levels. Venous blood was collected from these patients and the healthy individuals. One week after treatment, venous blood was collected once again in 60 patients with oral and maxillofacial squamous cell carcinoma (OSCC). RESULTS: The sCD44v6 concentration was not significantly different between patients with oral and maxillofacial malignancy and control group (P > 0.05). The levels of serum sCD44v6 in patients with OSCC and salivary carcinoma showed no difference with those in control group (P > 0.05). The sCD44v6 level in patients with stage III and IV disease was higher than that of patients with stage I and II and that of the control group, but the difference was not significant (P > 0.05). Serum sCD44v6 levels in patients with OSCC after treatment became lower than that prevailed during pretreatment (P < 0.05). CONCLUSION: The possible roles of CD44v6 in the diagnosis of oral and maxillofacial malignancy deserve further elucidation and evaluation. Serum sCD44v6 may be a valuable marker in monitoring treatment response in patients with OSCC.

Aberrant cytokine expression in serum of patients with adenoid cystic carcinoma and squamous cell carcinoma of the head and neck.

BACKGROUND: Squamous cell carcinoma (SCC) and adenoid cystic carcinoma (ACC) represent 2 clinically important subtypes of head and neck cancer. Our objective was to characterize and compare cytokine profiles in the systemic circulation of patients with SCC and ACC. METHODS: Multiplex analysis of 10 different cytokines (interleukin [IL]-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, granulocyte-macrophage colony-stimulating factor [GM-CSF], interferon [IFN]-gamma, and tumor necrosis factor [TNF]-alpha) in the serum of patients with SCC (n = 20) and ACC (n = 20) and healthy controls (n = 20) was performed using the Luminex fluorescent-bead technology. RESULTS: Patients with SCC as well as patients with ACC showed an altered cytokine profile compared with healthy individuals. In patients with SCC, significantly elevated serum levels of the proinflammatory cytokines, IL-6 and IL-8, were observed. In patients with ACC, IL-8 serum levels were significantly elevated, and IL-6 serum levels were only increased in a subset of patients. CONCLUSIONS: A similar serum cytokine profile, with the predominance of proinflammatory cytokines, was observed in patients with SCC and ACC. The newly defined cytokine profile in ACC patients may form the basis for future investigations to explore the role of cytokines in ACC tumor progression and their potential value as predictive biomarkers.

Detection of tumor DNA in plasma using whole genome amplification.

Altered microsatellite DNA in the blood of cancer patients may provide a novel means for tumor detection. Such alterations are a major characteristic of many types of tumor especially those associated with head or neck cancer. Moreover, recent evidence suggests that senescent tumor cells release DNA into the circulation, which is subsequently carried by the blood and thus enriched in the serum and plasma. We tested 10 head and neck cancer patients (5 with malignant melanomas (MM) and 5 with adenoid cystic carcinomas (ACC)) by polymerase chain reaction (PCR)-based microsatellite analysis of DNA from white blood cells and paired plasma samples. Our goal was to amplify two microsatellite markers, D1S243 and D19S246, which sometimes show microsatellite alterations in head and neck cancer patients. However amplification of fragments from three loci in the plasma samples proved impossible, probably due to the small amounts of DNA isolated. We used multiple displacement amplification (MDA) to amplify genomic DNA from the plasma samples. Two microsatellite fragments were amplified from whole genome amplified DNA. Among 5 heterozygote samples, 3 showed the same pattern in DNA samples from both blood cells and plasma but 2 showed loss of heterozygosity (LOH). Although further study is necessary to confirm whether the LOH found in this study reflects alteration in circulating tumor cell DNA, application of whole genome amplification may allow DNA analysis from limited amounts of such DNA and provide a minimally invasive diagnostic procedure and useful aid in therapy.

Expression of intercellular adhesion molecule (ICAM)-1 in adenoid cystic carcinoma of the head and neck.

OBJECTIVES/HYPOTHESIS: Adenoid cystic carcinoma of the head and neck (ACCHN) is characterized by late recurrence and frequent distant metastasis. Tumor attack by cytotoxic T lymphocytes and macrophages is mediated by the interaction of leukocyte function-associated antigen (LFA)-1 on lymphocytes with intercellular adhesion molecule (ICAM)-1 on the tumor surface. Thus, the reduced expression of ICAM-1 on tumor cells could contribute to their escape from host immune surveillance. To investigate the relationship between the clinical features of ACCHN and host immune surveillance, the expression of ICAM-1 and infiltration of T/natural killer (NK) cells and macrophages were immunohistochemically examined. STUDY DESIGN: Retrospective analysis of immunohistochemical tumor characteristics and clinical outcome. METHODS: Immunohistochemical study of ICAM-1, T/NK cells, and macrophages was performed on paraffin sections of 42 patients with ACCHN. The expression of T/NK cells and macrophages was represented by T-cell-restricted antigen (TIA)-1 and CD68 expression, respectively. The expression of these molecules and clinical features were analyzed. RESULTS: Of 42 ACCHN cases, 15, 9, and 15 patients were classified as ICAM-1 high, TIA-1 high, and CD68 high, respectively. The TIA-1 expression scores in ICAM-1-low patients were significantly lower than those in ICAM-1-high patients (1.3 +/- 3.7 vs. 8.3 +/- 12.7, P =.0031). The CD68 expression scores in ICAM-1-low patients were also significantly lower than those in ICAM-1-high patients (9.6 +/- 9.6 vs. 21.1 +/- 17.6, P =.0047). Moreover, ICAM-1-high patients had a significantly better disease-free survival rate (P =.043). CONCLUSIONS: Reduced expression of ICAM-1 may promote immune evasion and metastasis, resulting in poor prognosis in ACCHN.

Elevated carcinoembryonic antigen levels correlating with disease recurrence in a patient with adenoid cystic carcinoma.

BACKGROUND: Carcinoembryonic antigen (CEA) is an oncofetal glycoprotein involved in cell recognition and adhesion. Serum CEA has been extensively studied as a potential chemical marker for malignancy, most notably in patients with colon carcinoma. Serum CEA measurements have not been reported for patients with salivary gland carcinomas. METHODS: Serum CEA was measured in a case study using enzyme immunoassay with monoclonal antibody specific for CEA. Tissue was examined with standard histologic and immunohistologic methods. RESULTS: A patient was initially seen with adenoid cystic carcinoma (ACC) of the trachea and had a markedly elevated serum CEA level which declined after surgical resection. The serum CEA level became elevated again when the patient developed abdominal metastases and then declined after debulking of the tumor. Immunohistochemical study of the tumor was positive for CEA. CONCLUSIONS: The measurement of serum CEA levels may play a role in the management of patients with ACC. Clinical investigation utilizing monoclonal antibodies against CEA, for imaging and for the delivery of chemotherapy and radiotherapy may be worthwhile.

Association of IgA-anti-Fab autoantibodies with disease stage in head-and-neck cancer.

Patients with head-and-neck cancer commonly have immune defects. It was reported that these patients have raised serum IgA levels. We investigated whether IgA-anti-Fab autoantibodies, which occur in association with immune dysfunction, are present in patients with head-and-neck cancer. Sera of 101 patients with squamous-cell carcinoma (SCCHN) and 8 patients with adenoid cystic carcinoma (ACCHN) of the head and neck were tested in ELISA for IgA-anti-Fab autoantibody activity. IgA-anti-Fab serum activity was significantly higher in both SCCHN and ACCHN patients than in healthy controls. In patients with SCCHN, an association between disease stage and IgA-anti-Fab activity was established. Stage-IV patients had significantly higher IgA-anti-Fab than stage-I patients or healthy controls. Stage-II and stage-III patients had intermediate levels. Extremely high IgA-anti-Fab activity was observed in 7 patients who died within 6 months following testing, suggesting a relationship of autoimmunity with terminal disintegration of physiological body functions. IgA-anti-Fab autoantibodies may explain the occurrence of immune defects in patients with head-and-neck cancer.

[Significance of platelet aggregation in patients with malignant tumor of the head and neck].

Platelet aggregation test (PAgT) was done in 492 patients with malignant tumor of the head and neck, and in 100 healthy adults and 33 patients with benign tumors of the head and neck. The results showed: (1) The maximum aggregation rate (MAR) of the pretreatment group, recurring group and distant metastasis group of the malignant tumor patients were significantly higher than that of healthy adults, benign tumor and posttreatment group of malignant tumor patients; (2) There was no significant difference between MAR of healthy adults and the patients with benign tumor of the head and neck; (3) MAR was enhanced as the tumor progress. The Results indicate that (1) Platelet aggregation in malignant tumor of the head and neck is enhanced; and (2) PAgT might be used as a reference index for the nature, development, effect of treatment and prognosis of tumors. The mechanisms of enhancement of platelet aggregation in malignant tumor, and the role of platelet in pathogenesis of proliferation and metastasis of tumors were also discussed. The clinical use of the platelet-inhibitory agents for anti-proliferation and metastasis of tumors was in prospect.

Long-term intravenous hydroxyurea infusions in patients with advanced cancer. A phase I trial.

BACKGROUND: Hydroxyurea is an S-phase specific drug. Constant exposure of tumor cells with a low S-phase fraction to the agent may result in improved cell kill. Because of its short half-life, a continuous intravenous infusion may result in better tumor exposure than intake by mouth. The goal of this trial was to find the longest tolerable duration of a continued intravenous infusion of hydroxyurea (HU) given at escalating doses. METHODS: Eligible patients had histologically confirmed cancer without effective alternate therapy, normal blood counts, liver and kidney function. After giving informed consent, the infusion began via a permanent indwelling catheter utilizing a portable pump. Dose levels (in g/m2/d) were 0.5 for level I, 1.0 for level II, 1.66 for level III, and 2.5 for level IV. RESULTS: Fourteen patients were entered. Five were men. Median age was 56 years of age (range: 32-67), median performance status 1 (range: 0-2). Diagnoses were as follows: colorectal cancer, seven; unknown primary site, three; breast cancer, two; melanoma, one; and adenoid-cystic carcinoma, one. Nine patients were pretreated with chemotherapy. Three patients were entered per dose level, except on level I, were five were entered. The mean duration of infusion was 12 weeks on level I, 5 weeks on II, 3 on III, 1 on IV. Toxicity included leukopenia below 2.0 K/mm3 in one patient each on levels III and IV, thrombocytopenia below 100 K/mm3 in one patient each on levels II and IV, and stomatitis in three patients (one on level II and two on IV). This toxicity was dose limiting. One patient on level III, with an unknown primary, had an objective response. HU levels were measured by a modification of the Fabricius-Rajewsky method. Mean plasma levels in micrograms per milliliter (SEM) were as follows: level I, 3.6 (0.23); level II, 5.1 (0.57); level III, 10.1 (1.55); and level IV, 16.7 (one point). Fetal hemoglobin rose two-fold and five-fold in two patients on level I after 9 and 16 weeks on therapy, respectively. CONCLUSIONS: HU as a continuous intravenous infusion is well tolerated; the maximum duration of therapy is related inversely with the dose given. No major antitumor activity was seen. The greatest interest in the drug rests in its future use as a modulator and radiation potentiator. The increase in hemoglobin F was of interest and may be important in the treatment of sickle cell disease.

[The clinical entity of primary tracheal cylindroma (author's transl)]. / Zum klinischen Bild des primären Trachealzylindroms.

On the grounds of 5 own observations of primary tracheal cylindroma and under consideration of the cases mentioned in literature on this subject, the repeatingly occurring and, therefore, probably typical characteristics of this kind of tumour were described. In the course of the disease, three phases may quite easily be differentiated, and the special features of the main symptoms (dyspnoea, cough, expectoration, hoarseness) as well as apparently specific, however inconstant changes in the blood picture were set forth.