Oral Squamous Cell Carcinoma-Derived Cell-Free DNA Modulates Stemness and Migration of Oral Squamous Cell Carcinoma Cell Line by Inducing M2 Macrophage Polarization

ZHENG Shi-ze; MENG Lin; REN Fei-long; REN Chun-xia; LI Xing; WANG Dan-dan; SUN Hong-chen

doi:10.12182/20230560206

Volume 54 Issue 3

May 2023

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JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCES) > 2023 > 54(3): 510-516. > DOI: 10.12182/20230560206

ZHENG Shi-ze, MENG Lin, REN Fei-long, et al. Oral Squamous Cell Carcinoma-Derived Cell-Free DNA Modulates Stemness and Migration of Oral Squamous Cell Carcinoma Cell Line by Inducing M2 Macrophage Polarization[J]. Journal of Sichuan University (Medical Sciences), 2023, 54(3): 510-516. DOI: 10.12182/20230560206

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Oral Squamous Cell Carcinoma-Derived Cell-Free DNA Modulates Stemness and Migration of Oral Squamous Cell Carcinoma Cell Line by Inducing M2 Macrophage Polarization

ZHENG Shi-ze^{1, 2,},
MENG Lin^{1, 2},
REN Fei-long^{1, 2},
REN Chun-xia^{2, 3},
LI Xing^{1, 2},
WANG Dan-dan^{1, 2},
SUN Hong-chen^{1, 2, ,}

1.
Department of Pathology, Hospital of Stomatology, Jilin University, Changchun 130021, China
2.
Jilin Provincial Key Laboratory of Tooth Development, Jaw Bone Remodeling and Regeneration, Changchun 130021, China
3.
Department of Periodontology, Hospital of Stomatology, Jilin University, Changchun 130021, China

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Corresponding author:
SUN Hong-chen, E-mail: hcsun@mail.jlu.edu.cn
Received Date: October 12, 2022
Revised Date: April 24, 2023
Available Online: May 19, 2023
Published Date: May 19, 2023

Abstract

Abstract

Objective To investigate the effect of oral squamous cell carcinoma (OSCC)-derived cell-free DNA (cfDNA) on the polarization of macrophages and the regulatory effect of polarized macrophages on the stemness and migration of OSCC cells.
Methods A total of 30 OSCC tissue samples, 10 dysplastic oral tissue samples, and 10 normal oral tissue samples were collected. The status of all tissue samples was confirmed by pathology analysis. Immunohistochemical (IHC) staining and immunofluorescence (IF) staining were performed to examine the cell count and location of M2 macrophages in different types of oral tissue samples. The conditioned medium (CM) of OSCC cell line CAL-27 from the human tongue was collected and the cfDNA was concentrated and isolated for identification. The macrophages were treated by cfDNA and their morphological characteristics were observed under microscope. The expression levels of polarization-related indicators were determined by RT-qPCR. CAL-27 cell line was treated with macrophage CM induced by cfDNA and the expression levels of stemness-related genes were determined by RT-qPCR. Scratch-wound assay was conducted to verify that the migration ability of CAL-27 was modulated by macrophages induced by cfDNA.
Results There were more M2 macrophages in the deep connective tissue of dysplastic oral epithelium and the stroma of OSCC compared with those in the normal oral tissues (P<0.05). OSCC cell line CAL-27 could secret cfDNA of 10000-15000 bp in length. cfDNA secreted by CAL-27 could induced in macrophages significantly higher expression of M2-macrophage-related genes (P<0.05). cfDNA-treated macrophages induced significantly increased expression of stemness-related genes in CAL-27 cell line (P<0.05) and promoted the migration ability of CAL-27 cell line (P<0.05).
Conclusion OSCC-derived cfDNA promotes stemness and migration of OSCC cell line by inducing M2 macrophage polarization.
- Tumor-associated macrophage,
- Oral squamous cell carcinoma,
- Cell-free DNA,
- Polarization,
- Stemness

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References (31)

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