Elsevier

Lung Cancer

Volume 28, Issue 3, June 2000, Pages 225-235
Lung Cancer

Aneuploidy of chromosome 7 can be detected in invasive lung cancer and associated premalignant lesions of the lung by fluorescence in situ hybridisation

https://doi.org/10.1016/S0169-5002(00)00097-0Get rights and content

Abstract

In the present study the chromosomal status of seven invasive non small cell lung cancer specimens and associated premalignant lesions was investigated. By fluorescence in situ hybridisation (FISH) with centromere specific probes, an increase in the percentage of aneuploid cells from pre-invasive to invasive lesions could be demonstrated (mean 8.5 and 59%, respectively, for chromosome 7). Furthermore, mean chromosome copy numbers were higher in invasive carcinomas as compared to premalignant lesions, indicating polyploidization during tumor development. Increasing evidence suggests that aberrations of chromosome 7 occur early in the development of lung cancer. Whether these aberrations can be used as a biomarker for future neoplastic progression remains to be determined.

Introduction

Genetic alterations which characterize premalignant lesions of the lung are increasingly identified. Dysregulation of cell cycle regulatory genes and their respective gene products have been described [2], [3], [11], [12], [14], [19], [20], [23], [24], [28], [40], [55], [58] as well as mutations of the K-ras oncogene [5], [34], [47], [54] and allelic losses at chromosomal arms 3p and 9p [10], [13], [16], [17], [46], [51], [52] and 8p [56]. Furthermore, sequential molecular alterations have been suggested to occur during development to lung cancer, with deletions of 3p preceeding p53-gene alterations [4].

Distinct chromosomal aberrations have however, been rarely reported for preinvasive lesions of the lung. This may be due to difficulties in obtaining material suitable for conventional cytogenetic analysis as well as the low proliferative index of these lesions in vitro. In fact only three cases of successfully karyotyped dysplasias of the bronchial epithelium have been reported, with deletion of 17p13 or deletion of 3p as sole clonal abnormality [43], [44].

Numerical chromosomal alterations are a common finding in invasive lung cancer [37], [49], [50]. Polysomies of chromosome 7 were shown to be particularly frequent, occuring alone or in combination with other aberrations in 41% of non small cell lung cancer specimens [50]. Furthermore, aberrations of chromosome 7 could be demonstrated even in non neoplastic tissue adjacent to lung tumors [27] and in premalignant lesions of the upper aerodigestive tract. Polysomies of chromosomes 7 and 17 were detected with increasing frequency in normal epithelium, hyperplastic lesions and dysplastic lesions of the oral cavity [26]. Similar data were obtained for premalignant lesions and invasive cancer of the head and neck [42], [53]. Moreover, the presence of genetic aberrations in premalignant lesions appeared to be associated with progression to invasive cancer.

In the studies on preinvasive lesions of the head and neck, the technical limitations of conventional karyotypic analysis were overcome by interphase fluorescence in situ hybridisation (FISH) with chromosome specific probes, which provides the opportunity to evaluate cytogenetic aberrations on the single cell level. Moreover, since FISH can be performed on paraffin sections, chromosomal status can be related to distinct histological lesions.

In the present study, we therefore, used FISH with chromosome specific probes to investigate the chromosomal status of both invasive lung cancer and adjacent premalignant lesions. Chromosome 7 was targeted in all experiments. We aimed at demonstrating a clonal relationship of invasive and preinvasive lesions and thus determining aberrations which can be preferentially used in lung cancer screening procedures.

Section snippets

Materials and methods

Paraffin-sections from seven surgically resected non small cell lung cancer specimens (five adeno-carcinomas, two squamous cell carcinomas) were selected according to the concomitant presence of invasive carcinoma and premalignant lesions. The pathologic stage and grade of primary tumors is specified in Table 1. Premalignant and malignant lesions were identified on hematoxylin/eosin (H/E) stained deparaffinized sections and adjacent paraffin-sections were used in the FISH experiments. The

Results

All seven invasive carcinoma specimens were successfully investigated by FISH with centromeric probes. However, due to tissue artifacts, enumeration of signals was not possible in two of the seven premalignant lesions (Table 1).

Aneuploidy of chromosome 7 was detected in 100% (7/7) primary tumors. The percentage of aneuploid cells ranged from 10–94% (mean 59%). Premalignant lesions exhibited abnormalities of chromosome 7 in 80% (4/5) (range of aneuploid cells 5–21%; mean 8.5%; Fig. 4). Other

Discussion

Neoplasms of the upper aerodigestive tract and lung arise when genetic damage accumulates in a multistep process of malignant transformation, e.g. due to exposure to inhaled carcinogens. Chromosome 7 was shown to be an early marker of tumorigenesis in case of head and neck cancer, where polysomies of this chromosome have been demonstrated in hyperplastic and dysplastic epithelium by fluorescence in situ hybridisation [26], [42], [53]. Similar data for lung cancer are lacking, although Crowell

Conclusion

In the present study we demonstrated numerical aberrations of chromosome 7 to be present in invasive lung cancer and adjacent premalignant lesions of the lung. In contrast, we did not detect aneuploidy of chromosome 7 by FISH in normal bronchial mucosa /alveolar cells of lung cancer patients. These results suggest that alterations of chromosome 7 accompany the transition from normal lung tissue to hyperplastic lesions and may indicate a clonal relationship between primary tumor and associated

Acknowledgements

This study was supported by a grant from Kommission Onkologie.

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