Absence of mutations in the VHL gene but frequent loss of heterozygosity at 3p25 /26 in non-small cell lung carcinomas S. Miyakis a,c , T. Liloglou a,b , S. Kearney b , G. Xinarianos a,b , D.A. Spandidos c , J.K. Field a,b, * a Roy Castle International Centre for Lung Cancer Research, Liverpool L3 9TA, UK b Department of Clinical Dental Sciences, University of Liverpool, Liverpool L69 3BX, UK c Laboratory of Virology, Medical School, University of Crete, Heraklion 71409, Crete, Greece Received 16 July 2002; received in revised form 22 October 2002; accepted 4 November 2002 Abstract In this study we have examined 79 primary non-small cell lung tumours for the presence of mutations of the VHL gene as well as for allelic imbalance at the gene surrounding loci. While allelic imbalance was found in 83% of specimens, frequently affecting the whole 3p25  /p26 region, no mutations were detected in the VHL coding region. The fractional regional loss (FRL) was significantly higher in squamous cell carcinomas (0.746) than adenocarcinomas (0.493) (Wilcoxon P /0.002). This is the first investigation of the VHL gene mutational status in primary lung tumours. Our results indicate that mutation is not a common means of VHL inactivation in NSCLC. # 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: VHL; Non-small cell lung cancer; Single-strand conformational polymorphism; Heteroduplex analysis; Loss of heterozygosity; 3p 1. Introduction The VHL tumour suppressor gene, located on 3p25  / 26, is responsible for the Von Hippel /Lindau (VHL) hereditary cancer syndrome [1,2]. The VHL gene product is 213-amino acid protein with molecular weight of 24  /30 kDa [3]. Inactivation of VHL is possibly related to the promotion of angiogenesis. Loss of VHL protein (pVHL) function has been correlated with increased levels of hypoxia-inducible factor-1, which in turn, binds to genes responsible for the production of angiogenetic factors [4]. Somatic mutations of the gene and allelic loss at the VHL locus have also been detected in tumour types that do not participate in the homon- ymous disease [5,6]. VHL mutations have been reported in human lung cancer cell lines [7]. In particular, Sekido et al have studied 72 cell lines of SCLC and NSCLC origin and found only five mutations. However, no such data is available to date for primary lung carcinomas. The short arm of chromosome 3 is one of the most frequent areas of loss of heterozygosity (LOH) in human non-small cell lung cancer [8,9]. Allelic loss at one or more 3p regions has been detected in up to 100% of squamous cell carcinomas and 90% of adenocarcinomas of the lung [10]. LOH usually occurs at multiple sites on 3p, with frequencies exceeding 50% of cases for each site [9  /11]. In the present study we investigated the frequency of VHL genomic alterations in NSCLC and their possible association with the development and progression of the disease. We thus screened for muta- tions in the VHL coding region and splice sites and focused our allelic imbalance investigation on the loci surrounding the VHL gene at 3p25  /p26. Abbreviations: HA, heteroduplex analysis; LOH, loss of heterozygosity; MA, microsatellite alteration; NSCLC, non-small cell lung cancer; SqCCL, squamous cell carcinoma of the lung; SSCP, single strand conformational polymorphism; VHL, Von Hippel  / Lindau. * Corresponding author. Tel.:  /44-151-794-8900; fax:  /44-151- 794-8989 E-mail address: j.k.field@liv.ac.uk (J.K. Field). Lung Cancer 39 (2003) 273  /277 www.elsevier.com/locate/lungcan 0169-5002/02/$ - see front matter # 2002 Elsevier Science Ireland Ltd. All rights reserved. PII:S0169-5002(02)00506-8