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Abstract Hyperplastic candidiasis is characterized by thickening of the mucosal epithelia with Candida albicans infection with occasional progression to squamous cell carcinoma (SCC). Albicans is a critical factor in tumor development; however, the oncogenic mechanism is unclear. We have previously produced an animal model for hyperplastic candidiasis in the rat forestomach. In the present study, we investigate whether impaired DNA methylation and associated protein expression of tumor suppressor and DNA repair genes are involved in the SCC carcinogenesis process using this hyperplastic candidiasis model. Promoter methylation and protein expression were analyzed by methylation specific PCR and immunohistochemical staining, respectively, of 5 areas in the forestomachs of alloxan-induced diabetic rats with hyperplastic candidiasis: normal squamous epithelia, squamous hyperplasia, squamous hyperplasia adjacent to SCC, squamous hyperplasia transitioning to SCC, and SCC.

We observed nuclear p16 overexpression despite increases in p16 gene promoter methylation during the carcinogenic process. TIMP3 and RAR-β2 promoter methylation progressed until the precancerous stage but disappeared upon malignant transformation. In comparison, TIMP3 protein expression was suppressed during carcinogenesis and RAR-β2 expression was attenuated in the cytoplasm but enhanced in nuclei. ERCC1 and BRCA1 promoters were not methylated at any stage; however, their protein expression disappeared beginning at hyperplasia and nuclear protein re-expression in SCC was observed only for ERCC1. These results suggest that aberrant p16, RAR-β2, TIMP3, ERCC1, and BRCA1 expression might occur that is inconsistent with the respective gene promoter methylation status, and that this overexpression might serve to promote the inflammatory carcinogenesis caused by C. Albicans infection.

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Citation: Terayama Y, Matsuura T, Ozaki K (2016) Lack of Correlation between Aberrant p16, RAR-β2, TIMP3, ERCC1, and BRCA1 Protein Expression and Promoter Methylation in Squamous Cell Carcinoma Accompanying Candida albicans-Induced Inflammation. PLoS ONE 11(7): e0159090. Editor: Javier S. Castresana, University of Navarra, SPAIN Received: April 1, 2016; Accepted: June 27, 2016; Published: July 13, 2016 Copyright: © 2016 Terayama et al. This is an open access article distributed under the terms of the, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability: All relevant data are within the paper and its Supporting Information files.

Funding: The authors received no specific funding for this work. Competing interests: The authors have declared that no competing interests exist. Introduction Candida albicans ( C. Albicans) is known to induce oral, esophageal, and vaginal candidiasis [].

Severe Candida infections in patients are often associated with neoplastic disease [, ]. In addition, oral mucosa with hyperplastic candidiasis can become dysplastic from the hyperplasia and ultimately progress to carcinoma [,, ]. Albicans has the ability to produce carcinogens such as nitrosamines; thus, the persistent infection of C. Albicans in vivo acts as a promoter of oncogenesis [, ]. Therefore, C. Albicans infection serves as a critical factor in tumor development as well as a cause of inflammation. However, little is currently known concerning the oncogenic mechanism of C.

Albicans infection. Previously, we have reported that alloxan-induced diabetic rats frequently developed severe mucosal proliferative lesions with C. Albicans and bacterial infection in the forestomach and that these lesions progressed to squamous cell carcinoma (SCC) [, ]. Additionally, we have succeeded in inducing early onset proliferative and inflammatory lesions by dosing C.

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