cyclin-d1 and Pharyngeal-Neoplasms

to explore proliferative potential of peripheral blood lymphocytes of Chornobyl clean-up workers and persons with malignant neoplasms of the oral cavity, oropharynx and laryngopharynx by level of expression of cyclin D1 and quantitative parameters of cell cycle.. A total of 294 men aged (58.47 ± 7.32) were surveyed, 215 of them were Chornobyl clean"up workers (1986-1987), exposed at the dose range 10.43-3623.31 mSv; 49 persons of the control group and 30persons with malignant neoplasms of the oral cavity, oropharynx and laryngopharynx at III, IVА and IVВ stages ofthe disease. The analysis of parameters of cell cycle and proliferative activity of peripheral blood (PB) lymphocyteswas performed using the flow cytometry. The evaluation of distribution of cells by G0/G1, S, G2/M cell cycle phaseswas done in vivo and in in vitro. Proliferative potential was analyzed by level of expression of cytoplasmic protein ofcyclin D1.. Proliferative potential of PB lymphocytes of Chornobyl clean"up workers and persons with malignant neo"plasms of the oral cavity, oropharynx and laryngopharynx was assessed. An increase in the level of spontaneousсyclin D1 expression and disturbance of сyclin D1-dependent regulation of cell cycle of PB lymphocytes after mito"gen activation were determined in the Chornobyl clean-up workers. An increase in pool of cells in the S" and G2/M"phases of cell cycle was detected, which characterizes high proliferative potential of PB lymphocytes. These changesare most pronounced in the subgroup of persons with a radiation dose of D > 500 mSv, and in persons with oncolo"gical pathology.. A positive linear dependence has been established between the radiation dose and the number of cellsin the S"phase of cell cycle in the subgroup of Chornobyl clean"up workers with a radiation dose of D > 500 mSv. The detected changes of cyclin D1-dependent regulation of cell cycle and proliferative status of lymphocytes depend on the radiation dose, can be a manifestation of genome instability and be a cause for risks of oncogenesis, in a remote period after radiation exposure.. Meta. Doslidyty osoblyvosti proliferatyvnogo potentsialu limfotsytiv peryferychnoï krovi uchasnykivlikvidatsiï naslidkiv avariï na ChAES ta osib, khvorykh na zloiakisni novoutvorennia porozhnyny rota, rotovoï ta gor"tannoï chastyn glotky, za rivnem ekspresiï tsyklinu D1 ta kil'kisnymy pokaznykamy klitynnogo tsyklu.Materialy i metody. Obstezheno 294 cholovikiv u vitsi (58,47 ± 7,32) rokiv, sered nykh: 215 uchasnykiv likvidatsiïnaslidkiv avariï (LNA) na ChAES 1986–1987 rr., dozy zovnishn'ogo oprominennia 10,43–3623,31 mZv; 49 osib kont"rol'noï grupy ta 30 khvorykh na zloiakisni novoutvorennia porozhnyny rota, rotovoï ta gortannoï chastyn glotky III,IVA ta IVV stadiĭ zakhvoriuvannia. Analiz parametriv klitynnogo tsyklu i proliferatyvnoï aktyvnosti limfotsytivperyferychnoï krovi (PK) provodyly metodom protochnoï tsytometriï. Otsiniuvaly rozpodil klityn po G0/G1", S" iG2/M"fazakh klitynnogo tsyklu v umovakh in vivo ta in vitro. Proliferatyvnyĭ potentsial analizuvaly za rivnemekspresiï tsytoplazmatychnogo bilka tsyklinu D1.Rezul'taty. Provedeno otsinku proliferatyvnogo potentsialu limfotsytiv PK uchasnykiv likvidatsiï naslidkivavariï na ChAES ta osib, khvorykh na zloiakisni novoutvorennia porozhnyny rota. V uchasnykiv LNA na ChAES vyznache"no pidvyshchennia rivnia spontannoï ekspresiï tsyklinu D1 ta porushennia tsyklin D1"zalezhnoï reguliatsiï klitynnogotsyklu limfotsytiv PK pislia aktyvatsiï mitogenom. Vyznacheno zbil'shennia pulu klityn u S" ta G2/M"fazakh klityn"nogo tsyklu, shcho kharakteryzuie vysokyĭ proliferatyvnyĭ potentsial limfotsytiv PK. Dani zminy naĭbil'sh vyra"zheni u pidgrupi osib, oprominenykh v dozi D > 500 mZv, ta u patsiientiv z onkologichnoiu patologiieiu. Vysnovky. Vstanovleno pozytyvnu liniĭnu zalezhnist' mizh dozoiu oprominennia ta kil'kistiu klityn S"fazyklitynnogo tsyklu u pidgrupi uchasnykiv LNA na ChAES, oprominenykh u dozakh D > 500 mZv. Vyiavleni zminy tsyklin D1"zalezhnoï reguliatsiï klitynnogo tsyklu ta proliferatyvnogo statusu limfotsytiv zalezhat' vid dozy oprominennia,mozhut' buty proiavom nestabil'nosti genomu i sprychyniaty ryzyky onkogenezu u viddalenomu periodi pislia op"rominennia.

Topics: Adult; Aged; Aged, 80 and over; Case-Control Studies; Cell Cycle; Chernobyl Nuclear Accident; Cyclin D1; Emergency Responders; Humans; Lymphocytes; Male; Middle Aged; Mouth; Neoplasms; Occupational Exposure; Pharyngeal Neoplasms; Radiation Dosage; Radiation Exposure; Radiation Injuries; Ukraine

Omethoate is a broad category of organophosphorous pesticides (OPs) and has toxic effects on human health under long-term, low-dose exposure. However, the role of omethoate in cancer development remains elusive. The incidence of global head and neck squamous cell carcinomas (HNSCC) has markedly increased in recent years. Thus, we examined whether omethoate induced the proliferation of FaDu cells (a cell line of HNSCC) and if so, what the underlying mechanism was. The study revealed that omethoate induced FaDu cell growth in a dose- and time-dependent manner. Omethoate stimulated FaDu cell proliferation was mainly due to enhancing the G1 to S phase transition by flow cytometry analysis. We also found that omethoate up-regulated cyclin D1, a key gene controlling the G1-S transition. Furthermore, we showed that omethoate was capable of activating the Akt/GSK-3β signaling pathway. Blockage of Akt by siRNA or small molecule inhibitor significantly suppressed omethoate-induced cyclin D1 expression and cell proliferation. Collectively, these findings demonstrated for the first time that omethoate could induce the pharyngeal cancer cell proliferation by activation of the Akt/GSK-3β/cyclin D1 signaling pathway.

Topics: Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Dimethoate; Glycogen Synthase Kinase 3 beta; Humans; Pesticides; Pharyngeal Neoplasms; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Signal Transduction

Analyzing chromosomal amplifications delivers valuable information for identification of oncogenes. For carcinomas of the oral cavity only few genes have been identified in amplified regions. The aim of this study was to search genes in amplified regions as possible biomarkers and targets for novel therapies.. DNA from 10 carcinomas of the floor of the oral cavity was examined using a 500K Array GeneChip (Affymetrix 6.0) to detect chromosomal losses, gains or amplifications. Suspicious alterations were validated on tissue microarrays using fluorescence in situ hybridization (FISH) with respective probes.. FISH-validation on tissue arrays confirmed PPFIA1 amplifications as one of the most frequent events (32.6%). High (10-20 signals) and low (<10 signals) amplification of PPFIA1 was found in 10.9% (5/46) and 21.7% (10/46) tumours, respectively. Fine mapping with overlapping FISH probes showed co-amplification of PPFIA1 and the Cyclin D1 gene which are approximately 600 kb apart from each other, likely in the same amplicon.. PPFIA1 was frequently co-amplified with the Cyclin D1 gene in oral carcinomas and could present a biomarker as well as a novel target for specific gene therapy. Further studies are necessary to investigate the role of PPFIA1 in development and pathogenesis of oral carcinomas.

Topics: Adaptor Proteins, Signal Transducing; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Chromosomes, Human, Pair 11; Cyclin D1; DNA Probes; DNA, Neoplasm; Female; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Laryngeal Neoplasms; Male; Mouth Neoplasms; Neoplasm Grading; Neoplasm Staging; Oligonucleotide Array Sequence Analysis; Pharyngeal Neoplasms; Tissue Array Analysis

Second primary tumors and recurrences are an important problem in patients with head and neck squamous cell carcinoma. The purpose of this study was to determine the genetic changes in tumor samples to improve knowledge of tumor progression.. Copy number changes of 37 genes were analyzed by multiplex ligation-dependent probe amplification (MLPA) in 36 primary tumors and their corresponding 21 second primary tumors and 15 recurrences.. CCND1 and EMS1 amplifications and gain of BCL2L1 were the most common genetic alterations in the primary tumor, second primary tumor, and recurrence samples. Gains of ERBB2 and PTPN1 were associated with recurrences.. Specific genetic profiles for each group have been found. Similarities between primary tumor and second primary tumor and dissimilarity between primary tumor and recurrence suggest that clinicopathological criteria do not always accurately differentiate these entities. Genetic profiling may aid in the diagnosis and prognosis of these difficult cases.

Topics: Adult; Aged; bcl-X Protein; Carcinoma, Squamous Cell; Cathepsin B; Cell Cycle Proteins; Cortactin; Cross-Sectional Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA-Binding Proteins; Female; Gene Expression Profiling; Humans; Interleukin-18; Lamin Type A; Laryngeal Neoplasms; Male; Membrane Proteins; Middle Aged; Neoplasm Recurrence, Local; Neoplasms, Second Primary; Nucleic Acid Amplification Techniques; Oncogene Proteins; Pharyngeal Neoplasms; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Receptor, ErbB-2; Retrospective Studies; RNA-Binding Proteins; Transcription Factors; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

Several signaling pathways are involved in the progression of squamous cell carcinoma. Among them, activated PI3K/Akt may result in NF-κB nuclear translocation, thus leading to the transcription of genes enrolled in cellular invasion and proliferation, such as cyclin D1. This study sought to evaluate the expression of pAkt, NF-κB and cyclin D1 proteins in head and neck squamous cell carcinoma cell lines and their respective in vitro-obtained invasive clones.. Squamous cell carcinoma cell lines originating from the tongue, pharynx and the metastatic lymph node were submitted to an in vitro invasion assay to select invasive clones. All experimental groups were submitted to immunofluorescence and Western blot assays. Statistical analysis was performed through a Student's t-test with a significance level of 5%.. The pAkt and NF-κB expression differed from cytoplasm and nucleus depending on the studied cell line. The invasive clone from the tongue presented a network-like structure of pAkt's cytoplasmic expression. This lineage as well as the invasive clone from pharynx also showed pAkt and NF-κB nuclear transportation. Significant pAkt and NF-κB increases were observed in the tongue and pharynx invasive clones. Cyclin D1 was detected in the nucleus of all studied cells and was significantly enhanced in the invasive clones from tongue and pharynx.. This study suggests the participation of pAkt, NF-κB and cyclin D1 in the invasion process of head and neck squamous cell carcinoma. Moreover, cytoplasmic pAkt network-like structure was probably related to cytoskeleton changes presented during invasion.

Topics: Active Transport, Cell Nucleus; Carcinoma, Squamous Cell; Cell Line, Tumor; Clone Cells; Cyclin D1; Cytoskeleton; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Neoplasm Invasiveness; NF-kappa B; Pharyngeal Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; Tongue Neoplasms; Up-Regulation

Alteration of expression of tumour suppressor genes and cell cycle regulators may be responsible for oral and pharyngeal cancer development. We have studied the expression of p53, pRb, cyclin D(1) and cdk4 in 53 cases of oral and pharyngeal squamous cell carcinomas using immunohistochemistry. Tumour expression of all nuclear proteins was scored according to the percentage of positive cancer nuclei. Positive p53 expression was detected in 27/53 (50.94%) cases. Lack of pRb immunostaining was observed in 39/53 (73.58%) cases. Overexpression of cyclin D(1) was shown in 21 (39.62%) tumours. The overexpression of cdk4 was detected in 43/53 (81.13%) cases. There was no significant association among these cell cycle regulatory proteins. This implies that the aberration of an important cell cycle regulator may be sufficient to disrupt regulatory mechanism in a manner favouring tumourigenesis. In summary, our results suggest that inappropriate expression of p53, pRb, cyclin D(1) or cdk4 is likely to contribute to the development of oral and pharyngeal cancers. The lack of pRb expression and/or overexpression of cdk4 play a crucial role in the development of this malignancy.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Immunohistochemistry; Male; Middle Aged; Mouth Neoplasms; Neoplasm Proteins; Pharyngeal Neoplasms; Retinoblastoma Protein; Tumor Suppressor Protein p53

Hypopharyngeal squamous cell carcinomas (HPC) has an extremely poor prognosis. Characteristics of cell lines of head and neck squamous cell carcinomas including HPC were studied by various methods, e.g., chemosensitivity test and the immunohistochemistry staining method, to determine whether this poor prognosis is due to the biological behavior of this cancer. An HPC cell line was found to be resistant to anti tumor drugs, i.e., PEP, MTX and CPM and moderately sensitive to CDDP, 5-FU and ADM. Thermoresistance to hyperthermatic treatment and weak expression of ICAM-1 on the HPC cell line were observed. DNA synthesis by the HPC cell line was induced by stimulation with a low concentration of EGF and the amount of EGFR on these HPC cells was very high. In addition, cyclinD1 overexpression was found in the HPC cell line. Based on the above findings, further analysis of hypopharyngeal carcinoma cells and the development of a new treatment modality to control tumor growth and metastatic factors influencing the poor outcome are necessary to improve the prognosis of this cancer.

Topics: Antineoplastic Agents; Carcinoma, Squamous Cell; Cyclin D1; Cyclins; DNA, Neoplasm; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; ErbB Receptors; Head and Neck Neoplasms; Humans; Hypopharynx; Intercellular Adhesion Molecule-1; Oncogene Proteins; Pharyngeal Neoplasms; Tumor Cells, Cultured

Gene amplifications in the q13 band of chromosome 11 are among the most frequent genetic alterations in head and neck squamous cell carcinomas. Previous studies have suggested that such amplification is a marker of aggressive tumor evolution. Their potential for predicting subclinical lymph node invasion or disease recurrence was investigated in a prospective series of 50 oral and oropharyngeal carcinomas. Cell DNA content was also measured in 32 tumors of this series. Gene amplifications affecting the 11q13 band were detected in 11 of 50 (20%) patients, a relatively low frequency in comparison with data reported previously for other carcinomas of the upper aerodigestive tract, especially hypopharyngeal carcinomas. These gene amplifications were preferentially associated with aneuploidy. Cervical lymph nodes of 26 clinically N0 (Tumor-Node-Metastasis staging) patients were surgically explored. The frequency of 11q13 amplifications was very similar in the presence or in the absence of histological invasion, 3 of 15 (20%) and 2 of 11 (18%), respectively. Thus, 11q13 amplifications do not appear to be a reliable marker for prediction of subclinical lymph-node invasion in oral and oropharyngeal carcinomas. The detection of 11q13 amplifications was also not associated with a higher risk of disease recurrence. These data suggest that not only the prevalence but also the prognostic significance of 11q13 amplifications varies between tumors at different sites in the upper aerodigestive tract.

Topics: Adult; Aged; Aged, 80 and over; Aneuploidy; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 11; Cyclin D1; Female; Flow Cytometry; Gene Amplification; Humans; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasm Recurrence, Local; Oncogenes; Pharyngeal Neoplasms; Prognosis; Prospective Studies; Risk