demecolcine and Carcinoma--Squamous-Cell

The genetic content of head and neck squamous cell carcinomas is ill defined. Spectral karyotyping (SKY) is a new technique that allows the simultaneous detection of all chromosomal translocations by labeling each individual chromosome with different fluorescent agents. In the current study we used SKY to analyze cell lines and a primary tumor derived from head and neck squamous cell carcinomas (HNSCC) to delineate recurrent translocations and breakpoints.. Spectral karyotyping analysis of head and neck cancer.. Two cell lines (MDA886 and MSK922) and one primary tumor in short-term culture were subjected to metaphase growth arrest with colcemide in their exponential growth phase and fixed onto glass slides. Painting probes for each of the autosomes and the sex chromosomes were generated from flow-sorted human chromosomes using sequence-independent DNA amplification. The probes were labeled using a polymerase chain reaction-based reaction and hybridized to metaphase preparations for 2 days at 37 degrees C. Biotinylated probes were detected using avidin Cy5 and digoxigenin-labeled probes with an anti-mouse digoxigenin antibody followed by goat anti-mouse antibody conjugated to Cy5.5. Chromosomes were counterstained with 4,6-diamino-2-phenyliodole (DAPI), and a minimum of five metaphases were captured and analyzed for each case. Breakpoints on the SKY-painted chromosomes were determined by comparison of corresponding DAPI banding.. Spectral karyotyping analysis revealed a complex pattern of chromosomal abnormalities. A total of 66 translocations were identified in the three cases, with one new recurrent translocation at (der(4)t(4;20)(q35;?)). Nine complex translocations, involving three or more chromosomes, were identified in these cases. Overall, 96 breakpoints were assigned to metaphase chromosomes and another 74 breakpoints could not be assigned. Breakpoints most commonly involved chromosomes in genetic rearrangements were 1, 3, 5, 8, 13, 16, and 17.. Spectral karyotyping analysis reveals the true complexity of chromosomal aberrations in cell lines derived from head and neck squamous cell carcinomas. The use of SKY, in combination with other techniques, may allow for a more complete assessment of the genetic abnormalities of head and neck cancers and serve as a starting point for gene identification.

Topics: Carcinoma, Squamous Cell; Chromosome Aberrations; Chromosome Disorders; Cytogenetics; Demecolcine; DNA Probes; Gene Rearrangement; Head and Neck Neoplasms; Humans; In Situ Hybridization, Fluorescence; Karyotyping; Metaphase; Nucleic Acid Hybridization; Polymerase Chain Reaction; Sensitivity and Specificity; Spectrometry, Fluorescence; Translocation, Genetic; Tumor Cells, Cultured

Oral squamous cell carcinomas are characterized by complex, often near-triploid karyotypes with structural and numerical variations superimposed on the initial clonal chromosomal alterations. We used immunohistochemistry combined with classical cytogenetic analysis and spectral karyotyping to investigate the chromosomal segregation defects in cultured oral squamous cell carcinoma cells. During division, these cells frequently exhibit lagging chromosomes at both metaphase and anaphase, suggesting defects in the mitotic apparatus or kinetochore. Dicentric anaphase chromatin bridges and structurally altered chromosomes with consistent long arms and variable short arms, as well as the presence of gene amplification, suggested the occurrence of breakage-fusion-bridge cycles. Some anaphase bridges were observed to persist into telophase, resulting in chromosomal exclusion from the reforming nucleus and micronucleus formation. Multipolar spindles were found to various degrees in the oral squamous cell carcinoma lines. In the multipolar spindles, the poles demonstrated different levels of chromosomal capture and alignment, indicating functional differences between the poles. Some spindle poles showed premature splitting of centrosomal material, a precursor to full separation of the microtubule organizing centers. These results indicate that some of the chromosomal instability observed within these cancer cells might be the result of cytoskeletal defects and breakage-fusion-bridge cycles.

Topics: Antigens, Nuclear; Carcinoma, Squamous Cell; Cell Cycle Proteins; Centromere; Chromosome Aberrations; Chromosome Breakage; Chromosome Painting; Cytoskeleton; Demecolcine; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Karyotyping; Mitosis; Mouth Neoplasms; Nuclear Matrix-Associated Proteins; Nuclear Proteins; Tubulin; Tumor Cells, Cultured

The authors report the results of clinical elaboration and testing of the technic of intraesophageal intratumor sarcolysin iontophoresis associated with radiotherapy in 40 inoperable patients with cancer of the thoracic esophagus. A total focal dose of irradiation was on the average 1/3 less compared with the routine one. Iontophoresis was performed by a specially designed electrode with 5-10 mA during 60 minutes. The iontophoresis procedure was followed by irradiation. The immediate good results of the treatment were twice as frequent as in control groups. The treatment proceeded with less pronounced radiation response. An average survival of patients while using chemical drugs intophoresis proved to be equal to control data that is likely to be due to the far-advanced stage of the disease in most patients. An average survival in patients with less tumor proliferation was found to be longer than in analogous control series.

Topics: Adult; Antineoplastic Agents; Carcinoma, Basal Cell; Carcinoma, Papillary; Carcinoma, Squamous Cell; Demecolcine; Drug Therapy, Combination; Esophageal Neoplasms; Evaluation Studies as Topic; Female; Humans; Iontophoresis; Male; Melphalan; Middle Aged; Radiotherapy Dosage; Time Factors

The literature concerning the use of metaphase inducing agents as clinical sensitisers to radiation is briefly reviewed, and five cases are reported, which suggest that under ordinary clinical conditions, these agents are not likely to be of value. These results accord with animal experiments and a possible reason is suggested.

Topics: Adenocarcinoma; Adult; Aged; Anus Neoplasms; Carcinoma, Squamous Cell; Demecolcine; Female; Humans; Lip Neoplasms; Male; Middle Aged; Mitotic Index; Neoplasms; Radiation-Sensitizing Agents; Skin Neoplasms; Stomach Neoplasms

Topics: Adenocarcinoma; Aneuploidy; Biopsy; Carcinoma, Squamous Cell; Cell Division; Cell Movement; Cell Transformation, Neoplastic; Demecolcine; Diploidy; Ear Neoplasms; Humans; Karyotyping; Lymphoma; Mitosis; Nose Neoplasms; Pharyngeal Neoplasms

Topics: Bronchial Neoplasms; Carcinoma; Carcinoma, Squamous Cell; Cell Division; Colchicine; Demecolcine; Drug Therapy; Female; Humans; Mediastinum; Neoplasm Metastasis; Neoplasms; Prognosis; Skin Neoplasms; Thoracic Diseases; Uterine Cervical Neoplasms

Topics: Anus Neoplasms; Bowen's Disease; Carcinoma; Carcinoma, Squamous Cell; Colchicine; Demecolcine; Humans; Male; Penile Neoplasms; Penis; Skin Neoplasms

Topics: Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Colchicine; Demecolcine; Dermatology; Humans; Skin Diseases; Skin Neoplasms