Polymorphism within the Cyclin Dl Gene Is Associated with Prognosis in Patients with Squamous Cell Carcinoma of the Head and Neck1

We have examined the correlation of a frequent A/G polymorphism within exon 4 of the cyclin Dl gene (CCNDJ) with genetic susceptibility and clinical outcome in 384 patients with squamous cell carcinoma (SCC) of the head and neck. CCNDJ genotype frequencies were similar in the cases and 191 controls. Furthermore, the CCND1 genotype was not associated with susceptibility to SCC of the larynx, pharynx, or oral cavity. The influence of the CCND1 genotype on clinical outcome was also assessed. We found no correlation between genotype and tumor size (T1-T4), the involvement of nodes at presentation, or patient age and gender. However, the distribution of CCNDJ genotypes in cases with poorly differentiated tumors was significantly different to that in patients with well-/moderately differentiated tumors (P = 0.016; X22 = 8.71). Homozygosity for CCNDJ*G (GG genotype) was associated with poorly differentiated tumors (G3). We used Cox’s proportional hazards model to investigate the influence of the CCNDJ genotype on disease-free interval. CCNDJ GG was associated with reduced disease-free interval [P = 0.001; hazard ratio (HR) = 2.95; 95% confidence interval (CI) = 1.54-5.63]. This reReceived 3/25/98; revised 6/24/98; accepted 7/14/98. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with I 8 U.S.C. Section 1734 solely to indicate this fact. I Supported from grants from the North Staffordshire Medical Institute, West Midlands Locally Organized Research Scheme. and Schering Health Care. 2 To whom requests for reprints should be addressed, at Center for Cell and Molecular Medicine, School of Postgraduate Medicine, North Staffordshire Hospital, Thornburrow Drive Stoke-on-Trent ST4 7QB, England. Phone: 44-1782-555226; Fax: 44-1782-747319. mained significant after correction for tumor differentiation (P = 0.013; HR = 2.34; 95% CI 1.2-4.6) and tumor stage (P = 0.005; HR 2.64; 95% CI 1.34-5.19). Analysis of the data from patients with tumors at different sites showed that the CCNDJ GG genotype was associated with reduced disease-free interval in laryngeal (P 0.004; HR 3.63; 95% CI = 1.44-8.83) and pharyngeal (P = 0.006; HR = 3.48; 95% CI = 1.43-8.46) tumors. This is the first report of an association between CCNDJ polymorphism and prognosis in SCC of the head and neck. These data show that the CCNDJ GG genotype is an independent prognostic indicator of disease-free interval and supports initial observations in non-small cell lung cancer, that polymorphism within CCNDJ influences tumor behavior. INTRODUCTION SCCHN3 comprises approximately 5% of newly diagnosed malignancies in Northern Europe and the United States. Annually, more than 500,000 new cases are registered worldwide, and the incidence of the disease is increasing (1, 2). Despite improvements in therapeutic and reconstructive modalities, the overall survival rate remains relatively poor (3). Although chronic consumption of tobacco and alcohol are recognized as critical risk factors, genetic factors are also important, though few genes associated with genetic susceptibility have been identified (1, 4-7). It is also unclear which genetic factors determine tumor behavior and, therefore, patient prognosis. Further clinical outcome can vary among patients with SCCHN from the same site, with comparable tumor stage, nodal status, and histological grade (8, 9). A frequent target in carcinogenesis is the deregulation of G3-S phase progression of the cell cycle. Transition through G1 into S phase is regulated by the CDKs, which catalyze the phosphorylation of the RB protein. CDK activity is positively controlled by growth-promoting cyclins D, E, and A and negatively regulated by CDK inhibitors. During tumorigenesis, genetic alterations to CDKs (e.g., CDK4) and the regulators of CDK activity (cyclin Dl, pl6’ 4) and to RB, occur frequently in a number of cancers including SCCHN (10). The protooncogene CCNDJ is often activated during malignant development. Under normal mitogenic stimulation, D-type cyclin expression (cyclin Dl-3) drives the cell through the restriction point in G1 into S phase (10, 1 1). In malignancy, activation of CCNDJ, leading to loss of control of gene expression, occurs 3 The abbreviations used are: SCCHN, squamous cell carcinoma of the head and neck; NSCLC, non-small cell lung cancer; CCNDI, cyclin Dl gene; CDK, cyclin-dependent kinase; HR. hazard ratio: CI, confidence interval. Research. on October 16, 2017. © 1998 American Association for Cancer clincancerres.aacrjournals.org Downloaded from 2412 Cyclin Dl Polymorphism in Head and Neck Cancer Table 1 Clinica 1 characte ristics of patients with SCCHN Cases Laryngeal 5CC (n = 259) Pharyngeal 5CC (n = 87) Oral cavity 5CC (ii = 38) Mean age ± SD (yrs) 61.94 ± 10.1 58.86 ± 10.7 59.29 ± 10.5 Gender Male 234 (90.3)” 68 (78.2) 25 (65.8) Female 25 (9.7) 19 (21.8) 13 (34.2) Smoking habits none 14(6.2) 8(11.1) 4(12.9) 1-10 cigarettes/day 14 (6.2) 9 (12.5) 2 (6.5) I 1-25 cigarettes/day 125 (55.6) 30 (41.7) 14 (45.2) >25 cigarettes/day 72 (32.0) 25 (34.7) 1 1 (35.5) Alcohol consumption none 28 (12.4) 9 (12.5) 8 (25.8) 100 g/day 32 (14.2) 24 (33.3) 7 (22.6) T-factor TI 89 (39.6) 6 (7.4) 14 (38.9) 12 55(24.4) 20(24.7) 10(27.8) 13 53(23.6) 17(21.0) 4(11.1) T4 28 (12.4) 38 (46.9) 8 (22.2) Differentiation GO/I 12(9.5) 1 (1.7) 1 (3.6) G2 94 (74.6) 33 (55.0) 19 (67.9) G3 20 (15.9) 28 (43.3) 8 (28.6) Nodes at presentation Negative 92 (73.6) 16 (27.1) 12 (41.4) Positive 33 (26.4) 43 (72.9) 17 (58.6) 1 Percentage values displayed in parentheses. through several mechanisms. These include clonal cytogenetic rearrangements of the chromosomal region 1 1q13, where CCNDJ is located. In some parathyroid adenomas, an inversion involving 1 1q13 and 1 lplS results in CCNDI expression coming under the control of the parathyroid hormone gene promoter (12). In B cell malignancies, including centrocytic B cell lymphomas, CCNDJ expression is up-regulated under the influence of the immunoglobulin heavy chain enhancer as a result of a reciprocal chromosomal translocation at the BCLJ breakpoint t(1 l;14: ql3:q32; Refs. 13 and 14). Amplification of CCNDJ is also described in a number of tumors including those of the head and neck, breast, lung, bladder, and liver (15-20). In SCCHN, CCNDJ amplification leading to protein overexpression correlates with poor prognosis, assessed as reduced 5-year and overall survival (16, 21). Other studies have shown CCNDJ amplification and protein overexpression to be significantly associated with poor prognosis in primary hypopharyngeal and laryngeal SCCs (22, 23). In contrast, cyclin DI overexpression has been shown to correlate with good prognosis in tumors from NSCLC patients (18). Thus, deregulation of the control of CCNDI expression is important not only in understanding the mechanisms of carcinogenesis but also as a putative indicator of