口腔医学研究

口腔医学研究 ›› 2025, Vol. 41 ›› Issue (5): 378-385.DOI: 10.13701/j.cnki.kqyxyj.2025.05.004

• 口腔颌面外科学研究 • 上一篇    下一篇

牙龈卟啉单胞菌通过NDK/ATP/P2X7影响口腔鳞状细胞癌恶性表型的实验研究

魏巍1,2, 谭小容1,2, 李慕秋1,2, 龚忠诚1,2*, 李晨曦1,2*   

  1. 1.新疆医科大学第一附属医院(附属口腔医院)口腔颌面肿瘤外科 新疆 乌鲁木齐 830054;
    2.新疆维吾尔自治区口腔医学研究所 新疆 乌鲁木齐 830054
  • 收稿日期:2024-12-27 出版日期:2025-05-28 发布日期:2025-05-26
  • 通讯作者: *龚忠诚,E-mail:gump0904@aliyun.com李晨曦,E-mail:lichenximed@163.com
  • 作者简介:魏巍(1996~ ),男,江苏徐州人,硕士在读,研究方向:口腔颌面部肿瘤基础和临床研究。
  • 基金资助:
    国家自然科学基金(编号:82360481)天山英才医药卫生领军人才(编号:TSYC202301A001)新疆医科大学第一附属医院创新团队(党字[2023]52号)新疆维吾尔自治区研究生科研创新项目(编号:XJ2024G176)

Effect of Porphyromonas Gingivalis on Migration and Invasion of Oral Squamous Cell Carcinoma through NDK/ATP/P2X7 Pathway

WEI Wei1,2, TAN Xiaorong1,2, LI Muqiu1,2, GONG Zhongcheng1,2*, LI Chenxi1,2*   

  1. 1. Department of Oral and Maxillofacial Oncology & Surgery, the First Affiliated Hospital of Xinjiang Medical University, School / Hospital of Stomatology Xinjiang Medical University, Urumqi 830054, China;
    2. Stomatological Research Institute of Xinjiang Uygur Autonomous Region, Urumqi 830054, China
  • Received:2024-12-27 Online:2025-05-28 Published:2025-05-26

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摘要： 目的: 探索牙龈卟啉单胞菌(Porphyromonas gingivalis, P.gingivalis, P.g)分泌的核苷二磷酸激酶(nucleoside diphosphate kinase, NDK)是否通过耗竭口腔鳞状细胞癌(oral squamous cell carcinoma, OSCC)细胞外三磷酸腺苷(Adenosine triphosphate, ATP)调控P2X7门控从而影响OSCC的恶性表型。方法: 收集2022年1月~2023年12月就诊于新疆医科大学第一附属医院(附属口腔医院)口腔颌面肿瘤外科30例患者临床资料和OSCC组织样本通过免疫组织化学染色检测OSCC组织中P.g与P2X7的表达。通过体外培养SCC 9和SCC 25细胞,培养P.g(W83型),敲除P.g(W83型)中的NDK(P.g-△NDK)后并培养,建立细胞/细菌的共培养模型(采用细胞免疫荧光观察共培养模型),加入或不加入ATP,运用CCK-8、细胞划痕、Transwell、乳酸脱氧酶(lactate dehydrogenase,LDH)释放、检测P.g和P.g-△NDK对各组OSCC细胞增殖、迁移、侵袭能力及毒性的影响,并用蛋白质印迹实验验证其各组间P2X7蛋白的表达。结果: 在OSCC组织中P.g与P2X7的表达显著高于癌旁组织(P<0.0001);细胞学实验中,感染P.g的SCC9和SCC 25细胞增殖、迁移、侵袭能力增强(P<0.01),细胞毒性也增强(P<0.0001),感染P.g-△NDK后,SCC 9和SCC 25细胞增殖、迁移、侵袭能力进一步增强(P<0.01),细胞毒性减弱(P<0.0001) ;各组加入ATP后,Control组、感染P.g组及感染P.g-△NDK组的细胞增殖、迁移、侵袭能力均增强(P<0.05),细胞毒性均减弱(P<0.0001),其中感染P.g组上清液中ATP含量明显减少(P<0.05),P2X7表达与ATP含量呈正相关关系。结论: P.g分泌的NDK具有耗竭SCC 9和SCC 25细胞外ATP的能力,敲除NDK后,P2X7表达上调,从而增强其增殖、迁移、侵袭能力,并降低细胞毒性。

关键词: 牙龈卟啉单胞菌, 口腔鳞状细胞癌, 核苷二磷酸激酶, 三磷酸腺苷, 嘌呤能离子通道型受体7

Abstract: Objective: To explore the effect of nucleoside diphosphate kinase (NDK) secreted by Porphyromonas gingivalis (P.g) on proliferation, migration, and invasion of oral squamous cell carcinoma (OSCC) cells by hydrolyzing extracellular adenosine triphosphate (ATP), and to clarify the relationship between NDK and ATP on the impact of oral cancer. Methods: Clinical data and OSCC tissue samples from 30 patients who visited the Department of Maxillofacial Oncology Surgery at the Affiliated Stomatological Hospital of Xinjiang Medical University from January 2022 to December 2023 were collected. Immunohistochemical detection was used to determine the expression of P.g and P2X7 in oral squamous cell carcinoma tissues. By culturing human oral squamous cell carcinoma SCC 9 and SCC 25 cells in vitro with P.g (W83 type) or P.g knocking out NDK (P.g-△NDK), a cell/bacteria co-culture model was established (using cell immunofluorescence to observe the co-culture model). CCK-8, wound healing, Transwell, and LDH release were used to detect the effects of P.g and P.g-△NDK on proliferation, migration, invasion, and toxicity of human oral squamous cell carcinoma cells. The expression of P2X7 protein among the groups was verified using western blot. Results: The expressions of P.g and P2X7 were significantly higher in the tumor tissues than in the adjacent normal tissues (P<0.0001). The proliferation, migration, and invasion of SCC9 and SCC25 cells infected with P.g were enhanced (P<0.01), and their cytotoxicity was also enhanced (P<0.0001). After infection with P.g-△NDK, the proliferation, migration, and invasion of SCC9 and SCC25 cells were further enhanced (P<0.01), however, their cytotoxicity was weakened (P<0.0001). In all groups, the addition of ATP resulted in an enhancement of cell proliferation, migration, and invasion (P<0.05) and a weakening of cytotoxicity (P<0.0001). Among them, the ATP content in the supernatant of the P.g group was significantly reduced (P<0.05), and the expression of P2X7 was positively correlated with the ATP content. Conclusion: NDK secreted by P.g has the ability to deplete extracellular ATP of SCC 9 and SCC 25. After knocking out NDK, the expression of P2X7 is up-regulated, thereby enhancing its proliferation, migration, and invasion ability and reducing cytotoxicity.

Key words: Porphyromonas gingivalis, oral squamous cell carcinoma, nucleoside diphosphate kinase, adenosine triphosphate, purinergic ligand-gated ion channel 7