Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Novel Cytological Model for the Identification of Early Oral Cancer Diagnostic Markers: The Carcinoma Sequence Model

Version 1 : Received: 23 July 2021 / Approved: 26 July 2021 / Online: 26 July 2021 (08:19:05 CEST)

How to cite: Kawaharada, M.; Yamazaki, M.; Maruyama, S.; Abé, T.; Chan, N.N.; Kitano, T.; Kobayashi, T.; Maeda, T.; Tanuma, J. Novel Cytological Model for the Identification of Early Oral Cancer Diagnostic Markers: The Carcinoma Sequence Model. Preprints 2021, 2021070553. https://doi.org/10.20944/preprints202107.0553.v1 Kawaharada, M.; Yamazaki, M.; Maruyama, S.; Abé, T.; Chan, N.N.; Kitano, T.; Kobayashi, T.; Maeda, T.; Tanuma, J. Novel Cytological Model for the Identification of Early Oral Cancer Diagnostic Markers: The Carcinoma Sequence Model. Preprints 2021, 2021070553. https://doi.org/10.20944/preprints202107.0553.v1

Abstract

Most oral squamous cell carcinomas (OSCCs) arise from oral epithelial dysplasia; however, there is no useful marker for early OSCC detection, likely owing to the inability to continuously observe the carcinoma sequence. We aimed to establish an experimental model to observe changes in the sequential expression pattern of mRNA and protein in the same rat using liquid-based cytology techniques. Cytology specimens were collected from a 4-nitroquinoline 1-oxide-induced rat tongue cancer model at 2, 5, 8, 11, 14, 17, and 21 weeks. We examined candidate biomarker expression using immunocytochemistry and quantitative real-time PCR. The percentage of positively stained nuclei was calculated as the labeling index (LI). All rats had OSCC of the tongue at 21 weeks. Brd4 (Brd4), Myc (c-Myc), and Tp53 (p53) mRNA levels were upregulated during progression from negative for intraepithelial lesion or malignancy to SCC. Brd4- and c-Myc-LI were increased in low- and high-grade squamous intraepithelial lesions and SCC specimens. p53-LI was significantly increased in SCC specimens. Our experimental model allowed the observation of sequential morphological changes and mRNA and protein expression patterns in the same rat during carcinogenesis. By reducing the false negative rate, BRD4 and c-Myc can be useful markers for the early detection of OSCC.

Keywords

oral squamous cell carcinoma; oral cancer; Brd4; c-Myc; p53; cytology; liquid-based cytology; immunocytochemistry; 4-nitroquinoline 1-oxide

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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