Detection of chromosomal aberrations in cytologic brush specimens from head and neck squamous cell carcinoma

Abstract

METHODS. Brush specimens taken from the tumors of 20 patients with head and neck squamous cell carcinoma (HNSCC) and from the normal mucosa of 8 control 1 Department of Otorhinolaryngology and Head patients were analyzed by FISH using DNA probes for the chromosomes 1 and 7. and Neck Surgery, University Hospital Maas-The FISH results were compared with DNA flow cytometry and FISH results of the tricht and University of Maastricht, Maastricht, The Netherlands. solid tumor specimens. RESULTS. The results of this study showed that 15 of the 20 tumor brush specimens 2 Department of Molecular Cell Biology and Ge-contained numeric chromosomal aberrations in at least 5% of the cells collected. netics, University of Maastricht, Maastricht, The Chromosomal aberrations were detected in all brush specimens taken from tumors Netherlands. that were DNA aneuploid and showed aneusomy. The presence of these aberra-3 Department of Pathology, University Hospital tions correlated well with the classification ''suspicious for malignancy,'' which Maastricht, Maastricht, The Netherlands. was based on Papanicolaou stained slides of the same specimens. In the control group the percentage of chromosomally aberrant cells did not exceed 2%; in addition , no suspiciously malignant cells were observed in this group. CONCLUSIONS. This study reveals that the FISH technique can be applied diagnostically to brush specimens of HNSCC. The presence of chromosomal aberrations in ú 5% of the cells in these specimens can be considered as a marker for malignancy. KEYWORDS: fluorescence in situ hybridization, head and neck squamous cell carci-noma, cytology, DNA flow cytometry. T Presented in part at the 188 th Meeting of the he detection of genetic changes in the cytologic specimens of persons at high risk for the development of head and neck carcinoma in these persons. Epithelial cells can be obtained in a simple, noninva-sive manner by brushing the mucosa. Although exfoliate cytology The authors wish to thank Dr. E. J. M. Speel for using standard Papanicolaou staining is a valuable tool for the diagno-his useful advice on technical matters and for sis of several types of premalignancies and can be used in the head proofreading this article. and neck region for tumor diagnosis, the validity of the method de-Address for reprints: Joris A. Veltman, M.Sc., pends strongly on the representativity of the specimen and the experi-Department of Otorhinolaryngology and Head ence of the investigator. 1,2 Therefore, additional techniques have to and Neck Surgery, University Hospital Maas-be applied to obtain more objective diagnostic criteria. Fluorescence tricht, P.O. Box 5800, 6202 AZ Maastricht, The in situ hybridization (FISH) can be used to study the chromosomal Netherlands. constitution of cytologic specimens 3 and recently has been reported to be of use in the cytopathologic diagnosis of cervical neoplasia. 4,5

Clinical Implications of Human Papillomavirus in Head and Neck Cancers

“…For research purposes only, real-time PCR can be performed on microdissected tumor DNA normalized to a single-copy human gene to demonstrate one or more viral copies per tumor cell. 29,90,91 Possible future diagnostic tests that would likely have high specificity but low sensitivity for a diagnosis of HPV-associated HNSCC will include the detection of HPV16 DNA in plasma, 92 fluorescence ISH or ISH on papanicolou smears obtained directly from tumors, 6,93 and HPV16 E6 and E7 seroreactivity. 94,95 These are all active areas of investigation.…”

Noninvasive detection of aneuploid cells in laryngeal epithelial precursor lesions

“…A search of the literature disclosed only 1 study that used FISH analysis on brush sampling of laryngeal lesions. 17 That study indicated that most tumor brush specimens contained numeric chromosomal aberrations in at least 5% of the cells collected; however, the authors analyzed only 10 patients with laryngeal cancer. 17 To our knowledge, the current study is the first to use molecular cytogenetics as an aid in augmenting the clinical evaluation of laryngeal lesions using noninvasive brush sampling.…”

“…These results are in agreement with previous studies in which the most common chromosomal gains observed in LSCC were 8q gains and especially gains and amplifications of the MYC gene. 7,39,40 17,18,24 reported the presence of numerical aberrations for chromosomes 1 and 7 in dysplastic laryngeal lesions, especially in those that progressed to cancer. However, most of those studies used archival material (mainly paraffin-embedded tissue), and the use of paraffin-embedded tissue sections can be accompanied by confounding problems, such as nuclear slicing, variations of hybridization efficiency, and counting error, which may be responsible for the excess of monosomic cells.…”

“…17 That study indicated that most tumor brush specimens contained numeric chromosomal aberrations in at least 5% of the cells collected; however, the authors analyzed only 10 patients with laryngeal cancer. 17 To our knowledge, the current study is the first to use molecular cytogenetics as an aid in augmenting the clinical evaluation of laryngeal lesions using noninvasive brush sampling. The search for a cutoff point revealed that the percentage of patients with >4% ACs increased in relation to the histologic degree of dysplasia (36% in Group 3, 73% in Group 2, and 80% in Group 1; P ¼ .028); and both MYC and chromosome 8 differed significantly between the groups (P ¼ .030 and P ¼ .037, respectively).…”

“…[14][15][16] DNA ploidy has been analyzed by several groups, and the results indicate that aneuploidy is an early event in laryngeal carcinogenesis. [17][18][19] Studies using cytogenetic analysis, fluorescence in situ hybridization (FISH), and comparative genomic hybridization have demonstrated that a consistent set of chromosome regions frequently is altered in laryngeal SCC (LSCC), including gains on 3q, 5p, 8q, and 11q13 and losses on 3p and 9p. However, those studies either used archival material or were conducted on a limited number of patients with laryngeal cancer.…”