Persistent Exposure to Porphyromonas gingivalis Promotes Proliferative and Invasion Capabilities, and Tumorigenic Properties of Human Immortalized Oral Epithelial Cells.
Persistent Exposure to Porphyromonas gingivalis Promotes Proliferative and Invasion Capabilities, and Tumorigenic Properties of Human Immortalized Oral Epithelial Cells.
复制标题
持续暴露于牙龈卟啉单胞菌可促进人永生化口腔上皮细胞的增殖和侵袭能力以及致瘤特性
DOI：
10.3389/fcimb.2017.00057
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发表时间：
2017
影响因子：
5.7
通讯作者：
Pan Y
中科院分区：
文献类型：
--
作者：
Geng F;Liu J;Guo Y;Li C;Wang H;Wang H;Zhao H;Pan Y
近期的流行病学研究表明，口腔鳞状细胞癌（OSCC）与牙龈卟啉单胞菌（Porphyromonas gingivalis）之间存在显著关联，牙龈卟啉单胞菌是牙周病的主要致病菌。作为牙周炎的关键致病菌，牙龈卟啉单胞菌不仅会损害局部牙周组织，还能逃避宿主免疫系统，最终影响全身健康。然而，其在口腔鳞状细胞癌中的作用尚未明确。为探究牙龈卟啉单胞菌慢性感染对口腔鳞状细胞癌的潜在影响，并确定相关生物标志物作为有前景的治疗和预防靶点，我们建立了一种新模型，将人永生化口腔上皮细胞（HIOECs）以低感染复数（MOI）暴露于牙龈卟啉单胞菌中5 - 23周。通过增殖、伤口愈合、Transwell侵袭和明胶酶谱分析等实验，监测牙龈卟啉单胞菌感染的HIOECs的肿瘤生物学改变。对感染牙龈卟啉单胞菌15周的HIOECs进行微阵列和蛋白质组学分析，并通过对感染15周和23周的细胞进行定量实时聚合酶链反应（qRT - PCR）和（或）蛋白质免疫印迹法（western blot）验证部分选定数据。持续暴露于牙龈卟啉单胞菌会导致细胞形态改变，细胞周期中S期比例升高，增殖能力增强，并促进细胞迁移和侵袭特性。综合生物信息学分析和验证实验结果，NNMT、FLI1、GAS6、长链非编码RNA CCAT1、PDCD1LG2和CD274等肿瘤相关基因，可能被视为长期暴露于牙龈卟啉单胞菌后细胞发生肿瘤样转化的关键调节因子。此外，还发现了一些有用的临床生物标志物和新蛋白。总之，牙龈卟啉单胞菌可促进HIOECs的致瘤特性，这表明牙龈卟啉单胞菌慢性感染可能被视为口腔癌的潜在危险因素。从本模型中检测到的关键调节因子，可能用于监测伴有慢性牙周感染的口腔鳞状细胞癌的发展。
Recent epidemiological studies revealed a significant association between oral squamous cell carcinoma (OSCC) and Porphyromonas gingivalis, a major pathogen of periodontal disease. As a keystone pathogen of periodontitis, P. gingivalis is known not only to damage local periodontal tissues, but also to evade the host immune system and eventually affect systemic health. However, its role in OSCC has yet to be defined. To explore the underlying effect of chronic P. gingivalis infection on OSCC and to identify relevant biomarkers as promising targets for therapy and prevention, we established a novel model by exposing human immortalized oral epithelial cells (HIOECs) to P. gingivalis at a low multiplicity of infection (MOI) for 5–23 weeks. The P. gingivalis infected HIOECs were monitored for tumor biological alteration by proliferation, wound healing, transwell invasion, and gelatin zymography assays. Microarray and proteomic analyses were performed on HIOECs infected with P. gingivalis for 15 weeks, and some selected data were validated by quantitative real-time PCR and (or) western blot on cells infected for 15 and 23 weeks. Persistent exposure to P. gingivalis caused cell morphological changes, increased proliferation ability with higher S phase fraction in the cell cycle, and promoted cell migratory and invasive properties. In combining results of bioinformatics analyses and validation assays, tumor-related genes such as NNMT, FLI1, GAS6, lncRNA CCAT1, PDCD1LG2, and CD274 may be considered as the key regulators in tumor-like transformation in response to long-time exposure of P. gingivalis. In addition, some useful clinical biomarkers and novel proteins were also presented. In conclusion, P. gingivalis could promote tumorigenic properties of HIOECs, indicating that chronic P. gingivalis infection may be considered as a potential risk factor for oral cancer. The key regulators detected from the present model might be used in monitoring the development of OSCC with chronic periodontal infection.