Human Cancer Biology Differential Genetic and Functional Markers of Second Neoplasias in Hodgkin's Disease Patients

Purpose: The mechanisms involved in the appearance of a second neoplasia in patients with Hodgkin's disease (HD) are probably related to the genomic damage induced by the treatments administered and its repair. Here, we searched for some constitutive molecular mechanisms that in a basal manner may influence the behavior of HD patients. Experimental Design:We aimed to evaluate with the Comet Assay whether baseline, induced, and unrepaired DNA damage differ between HD patients who did not develop a second neoplasia (HD-NST), HD patients who developed a second tumor (HD-ST), and healthy individuals; and to identify, through cDNA microarray hybridization, an expression signature of genes that could discriminate between the three groups. Results: Baseline, induced, and unrepaired DNA damage was higher in HD-ST than in HD-NST and higher in the second group than in healthy donors. The genomic approach revealed two sets of genes that discriminated between healthy subjects and patients and between the three sets of individuals.Hsp40, RAD50, TPMT, Rap2a, E2F2, EPHX2, TBX21, and BATF were validated by reverse transcription-PCR. Conclusions: Functional and genomic techniques revealed that alterations in cell cycle, repair, detoxification, and stress response pathways could be involved in the development of HD and in the occurrence of a primary second neoplasia in these patients. Both approaches may be useful as biological markers in the clinical setting. Hodgkin's disease (HD) is a lymphoid neoplasia identified two centuries ago (1). Its tumor component consists of Hodgkin's cells and Reed-Sternberg's cells. It is accepted that these cells originate in the B cells of the germinal center of lymph nodes (2–4). Epidemiologic and serologic studies have reported evidence of a relationship with the EBV (5). The specific association of the virus with the neoplastic cells, its absence in normal lymphocytes, and the clonal integration of the virus into the Hodgkin's cells and Reed-Sternberg's cells are indicative of a mechanistic role of EBV in the transformation of Hodgkin's cells and ReedSternberg's cells (6, 7). The treatment is based on combined chemotherapy plus radiotherapy for early stages and on chemotherapy of differing intensities, with radiotherapy as consolidation in some cases, for advanced stages (8). The frequency of second tumors in these patients is higher than the incidence of primary tumors in the general population, one of the reasons for the highermortality in treatedHD patients (9). The risk of appearance of a solid tumor depends on age at treatment and the treatments received. This increased risk has been attributed to the effect of irradiation (10). The risk of lung cancer is higher and is dose-dependent in HD patients treated with radiotherapy, with or without chemotherapy (11). The development of lung cancer is also associated with chemotherapy containing alkylating agents (12, 13). Mantle radiotherapy received at young ages is associated with a risk of breast cancer 15 years or more later (14). The most frequent secondmalignancy associated with chemotherapy is acute nonlymphocytic leukemia. The risk of alkylatingrelated acute nonlymphocytic leukemia increases with increasing cumulative doses (15, 16). The mechanisms involved in the appearance of a second primary tumor inHDpatients who have received radiotherapy, chemotherapy, or combined treatment are probably related to genomic damage. The treatments administered act as carcinogens and can alter the DNA structure and/or activities (17, 18). Patients treated for HD have an excess risk of non–Hodgkin's Authors' Affiliations: Departments of Medical Oncology and Radiation Oncology, Hospital Universitario Puerta de Hierro, Majadahonda; and Molecular Diagnostic Unit, Molecular Pathology Program, Spanish Cancer Research Center, Madrid, Spain Received 12/16/08; revised 3/6/09; accepted 3/27/09; published OnlineFirst 7/21/09. Grant Support: FIS: PI05/0657; CM: S-GEN/0266/2006, ISCIII-RETIC RD06/0020/0020; SAF2007-60214; and Fundación Mutua Madrileña. The costs of publicationof this articlewere defrayed in part by the paymentof page charges. This articlemust therefore be herebymarked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Note: Y. Lorenzo and M. Provencio contributed equally to this paper. Requests for reprints: Félix Bonilla and Gemma Domínguez, Department of Medical Oncology, Hospital Universitario Puerta de Hierro, C/Manuel de Falla, 1, 28220-Majadahonda, Madrid, Spain. Phone: 34-91-1917769; Fax: 34-91-1916806; E-mail: [email protected] and gdominguez. [email protected]. F 2009 American Association for Cancer Research. doi:10.1158/1078-0432.CCR-08-3224 4823 Clin Cancer Res 2009;15(15) August 1, 2009 www.aacrjournals.org Research. on April 16, 2017. © 2009 American Association for Cancer clincancerres.aacrjournals.org Downloaded from Published OnlineFirst July 21, 2009; DOI: 10.1158/1078-0432.CCR-08-3224