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Effect of cell density on the malignant biological behavior of breast cancer by altering the subcellular localization of ANXA2 and its clinical implications

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Abstract

Objective

To investigate the subcellular localization of ANXA2 in breast cancer of different cell densities in humans and its relationship with the clinicopathological features of patients. To investigate the differences in ANXA2 subcellular localization in MDA-MB-231 cells of different cell densities. To compare the proliferation, invasion, and migration ability of MDA-MB-231 cells under different ANXA2 subcellular localization.

Methods

Immunohistochemistry was applied to detect the subcellular localization of ANXA2 in tissue sections of 60 breast cancer patients, and the association with ANXA2 subcellular localization was verified in conjunction with cell density. To investigate the relationship between cell density and clinicopathological data of breast cancer patients. To establish high- and low-density models of MDA-MB-231 breast cancer cell lines and verify the subcellular localization of ANXA2 using immunofluorescence and observation under confocal microscopy. The proliferation, migration, and invasion ability of MDA-MB-231 cells under different subcellular localization of ANXA2 were detected and compared using CCK-8 assay and Transwell assay. After changing the subcellular localization of ANXA2 in high-density MDA-MB-231 cells with PY-60, changes in biological behaviors of the compared MDA-MB-231 cells were observed. Two different 4T1 cell lines with high and low densities were implanted subcutaneously in nude mice to observe the effects of different cell densities on tumor growth in nude mice.

Results

The clinical data showed that breast cancer with high cell density had higher T stage and higher TNM stage, and the cell density was positively correlated with breast cancer mass size. ANXA2 was mainly localized to the cell membrane when the cell density of breast cancer cells was high and to the cytoplasm when the cell density was low. The CCK-8 assay showed that the proliferation rate of MDA-MB-231 cells increased (P < 0.05) after shifting the subcellular localization of ANXA2 from the cell membrane to the cytoplasm. Transwell invasion assay and Transwell migration assay showed that the invasion and migration ability of MDA-MB-231 cells increased significantly after the subcellular localization of ANXA2 was transferred from the cell membrane to the cytoplasm (P < 0.05). The animal experiments showed that high-density breast cancer cells could promote the growth of subcutaneous tumors in nude mice relative to low-density breast cancer cells.

Conclusion

Cell density can regulate the subcellular localization of ANXA2, and changes in the subcellular localization of ANXA2 are accompanied by the changes in the biological behavior of breast cancer.

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Acknowledgements

The authors are particularly grateful to all the people who have given us help with our article.

Funding

No funding was received.

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Authors and Affiliations

  1. Department of Thyroid and Breast Surgery, Affiliated Hospital of North Sichuan Medical College, Nanchong, 637000, Sichuan, China

    Jingtai Chen, Xiaohan Su, Qiao Tan, Hongyu Pu, Linxing Zhang, Yuqing Kang, Xiaobo Zhao, Shuangqiang Qian, Lingmi Hou & Yanchun Gao

  2. Department of Thyroid and Breast Surgery, Guang’an City People’s Hospital, Guang’an, Sichuan, China

    Yunhui Tang

  3. Department of Pathology, Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan, China

    Wei Hou

  4. Basic Medical College, North Sichuan Medical College, Nanchong, Sichuan, China

    Shishan Deng

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  1. Jingtai Chen
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  2. Xiaohan Su
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  3. Qiao Tan
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  4. Hongyu Pu
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All authors read and approved the manuscript and agree to be accountable for all aspects of the research in ensuring that the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Corresponding authors

Correspondence to Shuangqiang Qian, Shishan Deng, Lingmi Hou or Yanchun Gao.

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Ethical approval

The study was approved by Ethical Committee of Affiliated Hospital of North Sichuan Medical College and conducted in accordance with the ethical standards.

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Because of the retrospective nature of the study, patient informed consent for inclusion was waived.

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Chen, J., Su, X., Tan, Q. et al. Effect of cell density on the malignant biological behavior of breast cancer by altering the subcellular localization of ANXA2 and its clinical implications. Clin Transl Oncol 24, 2136–2145 (2022). https://doi.org/10.1007/s12094-022-02865-0

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