国际肿瘤学杂志

国际肿瘤学杂志 ›› 2026, Vol. 53 ›› Issue (1): 16-23.doi: 10.3760/cma.j.cn371439-20250530-00002

• 论著 • 上一篇    下一篇

靶向CD59抑制口腔鳞状细胞癌细胞增殖、迁移并诱导凋亡的机制研究

杨心婷1, 马腾宇1, 关树龙2, 杨梅3, 姜宙4, 杨心茹5, 姜良乾6, 高美华7, 徐颖婕7(), 丛蓓蓓7()   

  1. 1滨州医学院口腔医学院，烟台 264003
    2青岛市市南区人民医院外科，青岛 266520
    3青岛大学附属青岛市口腔医院预防科，青岛 266001
    4青岛大学附属妇女儿童医院生殖科，青岛 266034
    5山东第一医科大学第一附属医院（山东省千佛山医院）肿瘤科，济南 250014
    6山东省临沂市人民医院口腔科，临沂 276000
    7青岛大学附属青岛市口腔医院中心实验室，青岛 266001
  • 收稿日期:2025-05-30 出版日期:2026-01-08 发布日期:2026-01-13
  • 通讯作者: 丛蓓蓓，Email： xinruo1986@163.com；
    徐颖婕，Email： xyjdywe@163.com
  • 基金资助:
    国家自然科学基金(81902783);青岛市医疗卫生重点学科建设项目;山东省医药卫生口腔内科学重点学科(青岛大学附属青岛市口腔医院)资助项目

Research on the mechanism of targeting CD59 to inhibit proliferation， migration， and induce apoptosis in oral squamous cell carcinoma

Yang Xinting1, Ma Tengyu1, Guan Shulong2, Yang Mei3, Jiang Zhou4, Yang Xinru5, Jiang Liangqian6, Gao Meihua7, Xu Yingjie7(), Cong Beibei7()   

  1. 1School of Stomatology， Binzhou Medical University， Yantai 264003， China
    2Department of Surgery， Shinan District People's Hospital of Qingdao， Qingdao 266520， China
    3Department of Prevention， Qingdao Stomatological Hospital Affiliated to Qingdao University， Qingdao 266001， China
    4Department of Reproduction， Women and Children's Hospital， Qingdao University， Qingdao 266034， China
    5Department of Oncology， First Affiliated Hospital of Shandong First Medical University （Shandong Provincial Qianfoshan Hospital）， Jinan 250014， China
    6Department of Stomatology， Linyi People's Hospital of Shandong Province， Linyi 276000， China
    7Central Laboratory， Qingdao Stomatological Hospital Affiliated to Qingdao University， Qingdao 266001， China
  • Received:2025-05-30 Online:2026-01-08 Published:2026-01-13
  • Supported by:
    National Natural Science Foundation of China(81902783);Qingdao Key Health Discipline Development Fund;Shandong Provincial Key Medical and Health Discipline of Oral Medicine(Qingdao Stomatological Hospital Affiliated to Qingdao University)

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摘要：

目的 探讨靶向CD59对口腔鳞状细胞癌（OSCC）细胞增殖、迁移和凋亡的影响及其机制。方法 选取2020年9月至2022年9月青岛大学附属青岛市口腔医院收治的30例OSCC患者为研究对象，采用免疫组织化学法检测其肿瘤组织与配对的癌旁正常组织中CD59的表达情况。使用慢病毒转染的方式沉默、过表达人正常口腔角质细胞HOK、人舌鳞状细胞癌细胞SCC-9中CD59的表达，将细胞分为沉默CD59阴性对照组（siCD59-C组）、沉默CD59组（siCD59组）、过表达CD59组（CD59组）和过表达CD59阴性对照组（CD59-C组）。采用流式细胞术检测细胞凋亡，克隆形成实验、CCK-8法检测细胞增殖，伤口愈合实验检测细胞迁移，蛋白质印迹法检测PI3K、Akt、mTOR蛋白磷酸化水平。结果 免疫组织化学结果显示，CD59的表达在OSCC组织中（334.06±72.50）显著高于正常组织（125.77±56.60；t=3.20，P=0.033）。流式细胞术结果显示，siCD59-C组和siCD59组HOK细胞凋亡率分别为5.67%±0.83%、6.92%±0.51%，差异无统计学意义（t=2.23，P=0.089）；与siCD59-C组（17.79%±0.45%）相比，siCD59组SCC-9细胞凋亡率（38.03%±0.64%）显著增加（t=13.09，P<0.001）。CD59-C组和CD59组HOK细胞凋亡率分别为4.57%± 0.31%、5.25%±0.65%，差异无统计学意义（t=1.66，P=0.172）；与CD59-C组（18.19%±0.45%）相比，CD59组SCC-9细胞凋亡率（7.26%±0.28%）显著降低（t=35.76，P<0.001）。克隆形成实验结果显示，siCD59-C组和siCD59组HOK细胞克隆形成数分别为（350.53±6.31）、（367.01±7.36）个，差异无统计学意义（t=1.48，P=0.214）；与siCD59-C组[（418.46±6.75）个]相比，siCD59组[（326.02±5.15）个]SCC-9细胞克隆形成能力显著降低（t=3.46，P=0.026）。CCK-8法结果显示，CD59-C组和CD59组HOK细胞活力分别为2.72%±0.43%、2.60%±0.66%，差异无统计学意义（t=2.54，P=0.064）；与CD59-C组（1.21%±0.77%）相比，CD59组（2.09%± 0.45%）SCC-9细胞增殖能力显著提高（t=16.97，P<0.001）。伤口愈合实验结果显示，划痕24 h后，siCD59-C组和siCD59组HOK细胞迁移率分别为15.01%±1.56%、13.32%±2.08%，差异无统计学意义（t=1.25，P=0.279）；与siCD59-C组（17.67%±1.53%）相比，siCD59组（7.13%±1.22%）SCC-9细胞迁移能力显著下降（t=9.17，P<0.001）。蛋白质印迹法结果显示，siCD59-C组与siCD59组HOK细胞中PI3K、Akt、mTOR蛋白磷酸化水平差异均无统计学意义（均P>0.05）；与siCD59-C组相比，siCD59组SCC-9细胞中PI3K、Akt、mTOR蛋白磷酸化水平均明显降低（均P<0.001）。结论 CD59在OSCC组织中表达显著上调，沉默CD59可促进OSCC细胞凋亡，抑制其增殖和迁移，其机制可能与抑制PI3K/Akt/mTOR信号通路激活有关。

关键词: 口腔肿瘤, CD59抗原, 细胞凋亡, 细胞增殖, PI3K/Akt/mTOR信号通路

Abstract:

Objective To investigate the effects and mechanisms of targeting CD59 on the proliferation， migration， and apoptosis of oral squamous cell carcinoma （OSCC） cells. Methods Thirty patients with OSCC admitted to Qingdao Stomatological Hospital Affiliated to Qingdao University from September 2020 to September 2022 were selected as the research subjects. The expression of CD59 in their tumor tissues and paired adjacent normal tissues was detected by immunohistochemistry. CD59 expression was knocked down or overexpressed in HOK （normal oral keratinocytes） and SCC-9 （human tongue squamous cell carcinoma） cells via lentiviral transduction. Cells were divided into four groups： siCD59-C group （knockdown control）， siCD59 group （CD59 knockdown）， CD59 group （CD59 overexpression）， and CD59-C group （overexpression control）. Apoptosis was detected by flow cytometry； proliferation was evaluated via colony formation and CCK-8 assays； migration was examined by wound healing assay； and PI3K， Akt， and mTOR phosphorylation levels were detected by Western blotting. Results The immunohistochemical results showed that the expression of CD59 in OSCC tissues （334.06±72.50） was significantly higher than that in adjacent normal tissues （125.77±56.60； t=3.20， P=0.033）. The results of flow cytometry showed that the apoptosis rates of HOK cells in the siCD59-C group and the siCD59 group were 5.67%±0.83% and 6.92%±0.51%， respectively， with no statistically significant difference （t=2.23， P=0.089）. Compared with the siCD59-C group （17.79%±0.45%）， the apoptosis rate of SCC-9 cells in the siCD59 group （38.03%±0.64%） increased significantly （t=13.09， P<0.001）. The apoptosis rates of HOK cells in the CD59-C group and the CD59 group were 4.57%±0.31% and 5.25%±0.65%， respectively， with no statistically significant difference （t=1.66， P=0.172）. Compared with the CD59-C group （18.19%±0.45%）， the apoptosis rate of SCC-9 cells in the CD59 group （7.26%±0.28%） was significantly decreased （t=35.76， P<0.001）. The results of colony formation assay showed that the numbers of HOK cells clones formed in the siCD59-C group and the siCD59 group were 350.53±6.31 and 367.01±7.36， respectively， with no statistically significant difference （t=1.48， P=0.214）. Compared with the siCD59-C group （418.46±6.75）， the clone formation ability of SCC-9 cells in the siCD59 group （326.02±5.15） was significantly decreased （t=3.46， P=0.026）. The results of CCK-8 assay showed that the viability of HOK cells in the CD59-C group and the CD59 group were 2.72%±0.43% and 2.60%±0.66%， respectively， with no statistically significant difference （t=2.54， P=0.064）. Compared with the CD59-C group （1.21%±0.77%）， the proliferation ability of SCC-9 cells in the CD59 group （2.09%±0.45%） increased significantly （t=16.97， P<0.001）. The results of wound healing assay showed that 24 h after the scratch， the migration rates of HOK cells in the siCD59-C group and the siCD59 group were 15.01%±1.56% and 13.32%±2.08%， respectively， with no statistically significant difference （t=1.25， P=0.279）. Compared with the siCD59-C group （17.67%±1.53%）， the migration ability of SCC-9 cells in the siCD59 group （7.13%±1.22%） decreased significantly （t=9.17， P<0.001）. The results of Western blotting showed that there was no statistically significant difference in the phosphorylation levels of PI3K， Akt， and mTOR in HOK cells between the siCD59-C group and the siCD59 group （all P>0.05）. Compared with the siCD59-C group， the phosphorylation levels of PI3K， Akt， and mTOR in SCC-9 cells in the siCD59 group were significantly decreased （all P<0.001）. Conclusions CD59 expression is significantly upregulated in OSCC tissues. Knockdown of CD59 can promote apoptosis of OSCC cells， inhibit their proliferation and migration. The mechanism may be related to the inhibition of the activation of the PI3K/Akt/mTOR signaling pathway.

Key words: Mouth neoplasms, CD59 antigens, Apoptosis, Cell proliferation, PI3K/Akt/mTOR signaling pathway