Analysis of H3K27me3 expression and DNA methylation at CCGG sites in smoking and non-smoking patients with non-small cell lung cancer and their clinical significance

Zhu,Kunshou; Deng,Yujie; Weng,Guoxing; Hu,Dan; Huang,Cheng; Matsumoto,Keitaro; Nagayasu,Takeshi; Koji,Takehiko; Zheng,Xiongwei; Jiang,Wenhui; Lin,Gen; Cai,Yibin; Weng,Guibin; Chen,Xiaohui

doi:10.3892/ol.2018.8100

Analysis of H3K27me3 expression and DNA methylation at CCGG sites in smoking and non-smoking patients with non-small cell lung cancer and their clinical significance

Authors:
- Kunshou Zhu
- Yujie Deng
- Guoxing Weng
- Dan Hu
- Cheng Huang
- Keitaro Matsumoto
- Takeshi Nagayasu
- Takehiko Koji
- Xiongwei Zheng
- Wenhui Jiang
- Gen Lin
- Yibin Cai
- Guibin Weng
- Xiaohui Chen
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Affiliations: Department of Oncological Surgery, Fujian Cancer Hospital and Fujian Medical University Cancer Hospital, Fuzhou, Fujian 350014, P.R. China, Department of Chemotherapy, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China, Department of Cardiac Surgery, Fujian Provincial Hospital, Fuzhou, Fujian 350001, P.R. China, Department of Pathology, Fujian Cancer Hospital and Fujian Medical University Cancer Hospital Fuzhou, Fuzhou, Fujian 350014, P.R. China, Department of Medical Oncology, Fujian Cancer Hospital and Fujian Medical University Cancer Hospital Fuzhou, Fuzhou, Fujian 350014, P.R. China, Division of Surgical Oncology, Department of Translational Medical Sciences, Nagasaki University Graduate School of Biomedical Science, Nagasaki, Nagasaki 852‑8501, Japan, Department of Histology and Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Nagasaki 852‑8523, Japan
Published online on: February 21, 2018 https://doi.org/10.3892/ol.2018.8100
Pages: 6179-6188
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Smoking frequently leads to epigenetic alterations, including DNA methylation and histone modifications. The effect that smoking has on the DNA methylation levels at CCGG sites, the expression of trimethylation of histone H3 at lysine 27 (H3K27me3) and enhancer of zeste homolog 2 (EZH2), and their interactions in patients with non‑small cell lung cancer (NSCLC) were analyzed. There were a total of 42 patients with NSCLC, 22 with adenocarcinomas and 20 with squamous cell carcinomas enrolled in the present study. Expression of H3K27me3, EZH2 and proliferating cellular nuclear antigen (PCNA) were immunohistochemically detected. DNA methylation at CCGG sites was evaluated via histoendonuclease‑linked detection of DNA methylation sites. The apoptotic index of cancerous tissues obtained from patients of different smoking statuses was evaluated via the terminal deoxynucleotidyl‑transferase‑mediated dUTP‑biotin nick end labeling method. The association with clinicopathological data was calculated relative to different smoking statuses. Compared with the non‑smokers, smokers with NSCLC exhibited a significantly lower apoptotic index (P<0.05), and frequently had a lower level of DNA methylation at CCGG sites, lower H3K27me3 expression and a higher EZH2 expression (P<0.05). DNA methylation levels at CCGG sites were negatively correlated to the Brinkman index (P=0.017). Furthermore, there was a parallel association between the H3K27me3 and EZH2 expression levels in the majority of smokers, whereas in the majority of non‑smokers, there was a diverging association (P=0.015). There was a diverging association between the PCNA and EZH2 expression levels in the majority of smokers; however, in the majority of non‑smokers, there was a parallel association (P=0.048). In addition, the association between the CCGG methylation ratio and immunohistochemical expression of H3K27me3 was a parallel association in the majority of smokers, while in the majority of non‑smokers there was a diverging association (P=0.049). Conclusively, patients with NSCLC and different smoking statuses exhibit different epigenetic characteristics. Additionally, DNA methylation levels at the CCGG sites may have the ability to determine associations between the expression levels of H3K27me3, EZH2 and PCNA.

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