Volume 24 Issue 6
Jun.  2026
Turn off MathJax
Article Contents
Article Navigation >  Chinese Journal of General Practice  > 2026  >  24(6): 936-939
SHEN Yuqing, XU Rui, ZHENG Xianyu. Establishment of a stable PD-L1-overexpressing oral squamous carcinoma MOC-1 cell line and verification of its in vivo function[J]. Chinese Journal of General Practice, 2026, 24(6): 936-939. doi: 10.16766/j.cnki.issn.1674-4152.004525
Citation: SHEN Yuqing, XU Rui, ZHENG Xianyu. Establishment of a stable PD-L1-overexpressing oral squamous carcinoma MOC-1 cell line and verification of its in vivo function[J]. Chinese Journal of General Practice, 2026, 24(6): 936-939. doi: 10.16766/j.cnki.issn.1674-4152.004525
shu

Establishment of a stable PD-L1-overexpressing oral squamous carcinoma MOC-1 cell line and verification of its in vivo function

doi: 10.16766/j.cnki.issn.1674-4152.004525

  • College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, Hefei, Anhui 230032, China

Funds:

 2025AHGXZK40288

 AHWJ2023A20179

  • Received Date: 2025-12-11
    Available Online: 2026-11-10
    Abstract
  •   Objective  To construct a stable MOC-1 cell line with constitutive overexpression of programmed death-ligand 1 (PD-L1) and to investigate its effects on the in vivo biological behaviors of oral squamous cell carcinoma (OSCC).  Methods  The pCDH-PD-L1 overexpression plasmid was constructed via molecular cloning techniques, and then packaged into lentiviruses to infect murine oral squamous cell carcinoma (MOC-1) cells. Stable transfectants were selected with puromycin. PD-L1 expression at both the mRNA and protein levels was validated by RT-qPCR and Western blot analysis. Using a completely randomized design, 10 mice were divided into a PD-L1 overexpression group and a control group (5 mice per group) using a random number table. Stable cells were then subcutaneously implanted into mice to evaluate in vivo tumorigenicity. Immunohistochemistry (IHC) and immunofluorescence (IF) staining were performed to assess the expression of PD-L1 and Ki67, as well as the infiltration and cytotoxic function of CD8+ T cells in tumor tissues.  Results  The pCDH-PD-L1 recombinant plasmid was successfully constructed. RT-qPCR and Western blot analyses confirmed the successful establishment of a stable PD-L1-overexpressing MOC-1 cell line. In vivo subcutaneous tumor experiments showed that PD-L1 overexpression significantly accelerated tumor growth, resulting in markedly larger tumor volume and weight compared with the control group [(361.11±54.05) mm3 vs. (94.94±23.15) mm3, (14.0±4.9) g vs. (124.8±34.8) g, P < 0.001]. Immunological analyses indicated that the positive rate of PD-L1 [(60.5±4.2) % vs. (20.8±2.8) %, P < 0.001] and the proportion of Ki67-positive cells [(61.2±7.1) % vs. (30.4±5.5) %, P < 0.001] in tumor tissues of the overexpression group were both significantly increased. In contrast, the infiltration number of CD8+ T cells was significantly reduced [(5 023±961) cells/mm2 vs. (8 702±1 006) cells/mm2, P < 0.001], accompanied by impaired cytotoxic capacity.  Conclusion  We successfully established in vitro and in vivo models of PD-L1 overexpression based on the pCDH vector. PD-L1 overexpression significantly enhances the in vivo tumorigenicity of OSCC cells by inhibiting the infiltration and cytotoxic capacity of CD8+ T cells.

     

    • FullText(HTML)
  • loading
    • References(16)
  • [1]
    ALQUTUB S, ALQUTUB A, BAKHSHWIN A, et al. Histopathological predictors of lymph node metastasis in oral cavity squamous cell carcinoma: a systematic review and meta-analysis[J]. Front Oncol, 2024, 14: 1401211. DOI: 10.3389/fonc.2024.1401211.
    [2]
    XUE N N, WANG Y, WANG Z Y, et al. Trends in immunotherapy for oral squamous cell carcinoma[J]. Cell Oncol(Dordr), 2025, 48(5): 1159-1179. doi: 10.1007/s13402-025-01068-3
    [3]
    CHENG Y X, SONG Z Z, CHEN J, et al. Molecular basis, potential biomarkers, and future prospects of OSCC and PD-1/PD-L1 related immunotherapy methods[J]. Heliyon, 2024, 10(4): e25895. DOI: 10.1016/j.heliyon.2024.e25895.
    [4]
    KUJAN O, AGAG M, SMAGA M, et al. PD-1/PD-L1, Treg-related proteins, and tumour-infiltrating lymphocytes are associated with the development of oral squamous cell carcinoma[J]. Pathology, 2022, 54(4): 409-416. doi: 10.1016/j.pathol.2021.09.013
    [5]
    XU S B, WANG M Y, SHI X Z, et al. Influence of PD-1/PD-L1 on immune microenvironment in oral leukoplakia and oral squamous cell carcinoma[J]. Oral Dis, 2023, 29(8): 3268-3277. doi: 10.1111/odi.14332
    [6]
    TANG Q, CHEN Y, LI X J, et al. The role of PD-1/PD-L1 and application of immune-checkpoint inhibitors in human cancers[J]. Front Immunol, 2022, 13: 964442. DOI: 10.3389/fimmu.2022.964442.
    [7]
    KUJAN O, VAN SCHAIJIK B, FARAH C S. Immune checkpoint inhibitors in oral cavity squamous cell carcinoma and oral potentially malignant disorders: a systematic review[J]. Cancers, 2020, 12(7): 1937. DOI: 10.3390/cancers12071937.
    [8]
    MU H R, ZHANG Q, ZUO D Q, et al. Methionine intervention induces PD-L1 expression to enhance the immune checkpoint therapy response in MTAP-deleted osteosarcoma[J]. Cell Rep Med, 2025, 6(3): 101977. DOI: 10.1016/j.xcrm.2025.101977.
    [9]
    GAO Y, FENG Y F, LIU S C, et al. Immune-independent acquired resistance to PD-L1 antibody initiated by PD-L1 upregulation via PI3K/AKT signaling can be reversed by anlotinib[J]. Cancer Med, 2023, 12(14): 15337-15349. doi: 10.1002/cam4.6195
    [10]
    陈志红, 吴亚东. 程序性死亡受体-1及其配体在口腔鳞状细胞癌中的研究进展[J]. 华西口腔医学杂志, 2020, 38(4): 449-453.

    Chen Z H, Wu D Y. Development of programmed death receptor-1 and programmed death receptor-1 ligand in oral squamous cell carcinoma[J]. West China Journal of Stomatology, 2020, 38(4): 449-453.
    [11]
    ZHOU Y J, LI G L, WANG J Y, et al. PD-L1: expression regulation[J]. Blood Sci, 2023, 5(2): 77-91. doi: 10.1097/BS9.0000000000000149
    [12]
    YANG Z, LIU X P, ZHU J, et al. Inhibiting intracellular CD28 in cancer cells enhances antitumor immunity and overcomes anti-PD-1 resistance via targeting PD-L1[J]. Cancer Cell, 2025, 43(1): 86-102. e10. doi: 10.1016/j.ccell.2024.11.008
    [13]
    房恩岳, 张丽萍, 马雪征, 等. 慢病毒载体系统及其安全性研究进展[J]. 病毒学报, 2023, 39(4): 1181-1192.

    Fang E Y, Zhang L P, Ma X Z, et al. Research progress on lentiviral vector systems and their safety issue[J]. Chinese Journal of Virology, 2023, 39(4): 1181-1192.
    [14]
    GARCIA-DIAZ A, SHIN D S, MORENO B H, et al. Interferon receptor signaling pathways regulating PD-L1 and PD-L2 expression[J]. Cell Rep, 2017, 19(6): 1189-1201. doi: 10.1016/j.celrep.2017.04.031
    [15]
    JUNEJA V R, MCGUIRE K A, MANGUSO R T, et al. PD-L1 on tumor cells is sufficient for immune evasion in immunogenic tumors and inhibits CD8 T cell cytotoxicity[J]. J Exp Med, 2017, 214(4): 895-904. doi: 10.1084/jem.20160801
    [16]
    孙奥龙, 张丽柯, 徐全晓, 等. 非小细胞肺癌患者中GZMB和PD-L1的表达及两者与预后的关系[J]. 中华全科医学, 2024, 22(11): 1863-1867. doi: 10.16766/j.cnki.issn.1674-4152.003751

    Sun A L, Zhang L K, Xu Q X, et al. The expression of GZMB and PD-L1 and their relationship with prognosis in non-small cell lung patients[J]. Chinese Journal of General Practice, 2024, 22(11): 1863-1867. doi: 10.16766/j.cnki.issn.1674-4152.003751
    • Relative Articles
    • Supplements(0)
    • Cited By

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(4)  / Tables(1)

    Get Citation
    PDF
    XML

    Article Metrics

    Article views (8) PDF downloads(1) Cited by()
    Proportional views
    Related

    Address：No. 287, Changhuai Road, Bengbu, Anhui Province, 233004, P.R.ChinaphoneNo：0552-3066635； 0552-3051890Email：zhqkyx@163.com

    Copyright © Chinese Journal of General Practice皖ICP备2020018345号-2

    Supported by: Beijing Renhe Information Technology Co., Ltd.    

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return