Dynamic Succession of Urokinase-Type Plasminogen Activator in an Oral Squamous Cell Carcinoma Model

LUO Yuwen, HUANG Xuelin, CUI Bomiao, JI Ning, ZHANG Ping

Abstract

Objective 

To systematically characterizes the temporal changes in urokinase-type plasminogen activator (uPA) over the course of neoplastic progression using a mouse oral squamous cell carcinoma (OSCC) model induced by 4-nitroquinoline-1-oxide (4-NQO).

Methods 

A total of 65 wild-type C57BL/6 mice of 5 weeks old were randomly assigned to two groups, a 4-NQO group (n = 50), which received daily administration of 100 μg/mL 4-NQO in drinking water, and a control group (n = 15), which received sterile water. At 12, 16, 20, 22, and 24 weeks, 10 mice from the 4-NQO group and 3 from the control group were randomly selected, weighed, and sacrificed. Tongue tissues were collected for hematoxylin-eosin (HE) staining to preliminarily assess OSCC development, and for immunofluorescence staining and quantitative real-time PCR to evaluate dynamic uPA expression in tongue tissues during OSCC progression.

Results 

After 16 weeks of exposure, 4-NQO-treated mice exhibited significantly lower body mass compared with that of the controls (P < 0.05) and the weight loss became increasingly more pronounced over time. Histopathological changes in tongue tissues progressed in a clearly time-dependent manner—hyperplasia and mild dysplasia emerged at week 12, while moderate-to-severe dysplasia and carcinoma were observed by week 22, yielding a tumorigenic rate of 25%, which escalated to 70% by week 24. Immunofluorescence and qPCR analyses demonstrated a pronounced, progressive up-regulation of uPA expression in lesional tissues as OSCC progressed (P < 0.0001).

Conclusion 

This study not only confirmed the uniqueness of the 4-NQO model in OSCC research, but also revealed the changes in uPA during tumor invasion. These findings provide a theoretical foundation for the development of early diagnosis and precision treatment strategies, holding significant potential clinical value and research importance for improving patient prognosis.

 

Keywords: Oral squamous cell carcinoma, 4-nitroquinoline 1-oxide, Urokinase-type plasminogen activator


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References


SUNG H, FERLAY J, SIEGEL RL , et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2021, 71(3): 209-249. doi: 10. 3322/caac.21660.

SAIKIA P J, PATHAK L, MITRA S, et al. The emerging role of oral microbiota in oral cancer initiation, progression and stemness. Front Immunol, 2023, 14: 1198269. doi: 10.3389/fimmu.2023.1198269.

OIKAWA Y, MICHI Y, TSUSHIMA F, et al. Management of retropharyngeal lymph node metastasis in oral cancer. Oral Oncol, 2019, 99: 104471. doi: 10.1016/j.oraloncology.2019.104471.

LINSEN S S, GELLRICH N C, KRÜSKEMPER G. Age- and localization-dependent functional and psychosocial impairments and health related quality of life six months after OSCC therapy. Oral Oncol, 2018, 81: 61-68. doi: 10.1016/j.oraloncology.2018.04.011.

MEIER J K, SCHUDERER J G, ZEMAN F, et al. Health-related quality of life: a retrospective study on local vs. microvascular reconstruction in patients with oral cancer. BMC Oral Health, 2019, 19(1): 62. doi: 10.1186/s12903-019-0760-2.

TAN Y, WANG Z, XU M, et al. Oral squamous cell carcinomas: state of the field and emerging directions. Int J Oral Sci, 2023, 15(1): 44. doi: 10. 1038/s41368-023-00249-w.

NERLOV C, De CESARE D, PERGOLA F, et al. A regulatory element that mediates co-operation between a PEA3-AP-1 element and an AP-1 site is required for phorbol ester induction of urokinase enhancer activity in HepG2 hepatoma cells. EMBO J, 1992, 11(12): 4573-4582. doi: 10.1002/j. 1460-2075.1992.tb05559.x.

GAIRE M, MAGBANUA Z, MCDONNELL S, et al. Structure and expression of the human gene for the matrix metalloproteinase matrilysin. J Biol Chem, 1994, 269(3): 2032-2040. doi: 10.1016/S0021-9258(17)42131-4.

SATO H, SEIKI M. Regulatory mechanism of 92 kDa type Ⅳ collagenase gene expression which is associated with invasiveness of tumor cells. Oncogene, 1993, 8(2): 395-405.

MA Y, LIU Y, MENG H. Prognostic evaluation of oral squamous cell carcinoma based on pleiotrophin, urokinase plasminogen activator, and glycoprotein nonmetastatic melanoma protein B expression. Medicine (Baltimore), 2023, 102(45): e35634. doi: 10.1097/MD.0000000000035634.

DERYUGINA E I, QUIGLEY J P. Tumor angiogenesis: MMP-mediated induction of intravasation- and metastasis-sustaining neovasculature. Matrix Biol, 2015, 44-46: 94-112. doi: 10.1016/j.matbio.2015.04.004.

KANG Y, CHEN J, LI X, et al. Salivary KLK5 and uPA are potential biomarkers for malignant transformation of OLK and OLP. Cancer Biomark, 2021, 31(4): 317-328. doi: 10.3233/CBM-203105.

TRAN Q, MADDINENI S, ARNAUD E H, et al. Oral cavity cancer in young, non-smoking, and non-drinking patients: a contemporary review. Crit Rev Oncol Hematol, 2023, 190: 104112. doi: 10.1016/j.critrevonc. 2023.104112.

ZHANG Q, SHI J. Research progress in animal models of oral leukoplakia. J Oral Sci Res, 2025, 41(3): 179-183. doi: 10.13701/j.cnki. kqyxyj.2025.03.001.

SAGHEER S H, WHITAKER-MENEZES D, HAN J Y S, et al. 4NQO induced carcinogenesis: a mouse model for oral squamous cell carcinoma. Methods Cell Biol, 2021, 163: 93-111. doi: 10.1016/bs.mcb.2021.01.001.

HE D, ZHANG J, WU W, et al. A novel immunodeficient rat model supports human lung cancer xenografts. FASEB J, 2019, 33(1): 140-150. doi: 10.1096/fj.201800102RR.

LI Q, DONG H, YANG G, et al. Mouse tumor-bearing models as preclinical study platforms for oral squamous cell carcinoma. Front Oncol, 2020, 10: 212. doi: 10.3389/fonc.2020.00212.

KANOJIA D, VAIDYA M M. 4-nitroquinoline-1-oxide induced experimental oral carcinogenesis. Oral Oncol, 2006, 42(7): 655-667. doi: 10.1016/j.oraloncology.2005.10.013.

DOWNES D J, CHONOFSKY M, TAN K, et al. Characterization of the mutagenic spectrum of 4-nitroquinoline 1-oxide (4-NQO) in Aspergillus nidulans by whole genome sequencing. G3 (Bethesda), 2014, 4(12): 2483-2492. doi: 10.1534/g3.114.014712.

CORDANI M, BUTERA G, PACCHIANA R, et al. Mutant p53-associated molecular mechanisms of ROS regulation in cancer cells. Biomolecules, 2020, 10(3): 361. doi: 10.3390/biom10030361.

SEQUEIRA I, RASHID M, TOMÁS I M, et al. Genomic landscape and clonal architecture of mouse oral squamous cell carcinomas dictate tumour ecology. Nat Commun, 2020, 11(1): 5671. doi: 10.1038/s41467-020-19401-9.

LIU Y, MIAO K Y, HU Y H, et al. Correlation between p53 staining and TP53 mutation status in 60 patients with oral squamous cell carcinoma. China J Oral Maxillofac Surg, 2025, 23(3): 244-252. doi: 10.19438/j.cjoms. 2025.03.006.

CHEN Y F, LIU C J, LIN L H, et al. Establishing of mouse oral carcinoma cell lines derived from transgenic mice and their use as syngeneic tumorigenesis models. BMC Cancer, 2019, 19(1): 281. doi: 10.1186/s12885-019-5486-7.

LU G, WANG D, QIN X, et al. Detection and delineation of squamous neoplasia with hyperspectral imaging in a mouse model of tongue carcinogenesis. J Biophotonics, 2018, 11(3): 10.1002/jbio. 201700078. doi: 10.1002/jbio.201700078.

LEE Y M, HSU C L, CHEN Y H, et al. Genomic and transcriptomic landscape of an oral squamous cell carcinoma mouse model for immunotherapy. Cancer Immunol Res, 2023, 11(11): 1553-1567. doi: 10. 1158/2326-6066.CIR-23-0133.


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