Lung Adenocarcinoma Cells Promote Self-Migration and Self-Invasion by Activating Neutrophils to Upregulate Notch3 Expression of Cancer Cells

Weidong Peng; Youjing Sheng; Han Xiao; Yuanzi Ye; Louis Boafo Kwantwi; Lanqing Cheng; Yuanchong Wang; Jiegou Xu; Qiang Wu

doi:10.3389/fmolb.2021.762729

Lung Adenocarcinoma Cells Promote Self-Migration and Self-Invasion by Activating Neutrophils to Upregulate Notch3 Expression of Cancer Cells

Front Mol Biosci. 2022 Jan 18:8:762729. doi: 10.3389/fmolb.2021.762729. eCollection 2021.

Authors

Weidong Peng^{1

2}, Youjing Sheng^{1

2}, Han Xiao¹, Yuanzi Ye¹, Louis Boafo Kwantwi², Lanqing Cheng³, Yuanchong Wang⁴, Jiegou Xu⁵, Qiang Wu^{1

2}

Affiliations

¹ Department of Pathology, The First Affiliated Hospital of Anhui Medical University, Hefei, China.
² Department of Pathology, School of Basic Medical Science, Anhui Medical University, Hefei, China.
³ Department of Pathology, The Second Affiliated Hospital of Anhui Medical University, Hefei, China.
⁴ Department of Neonatology, Anhui Provincial Children's Hospital, Hefei, China.
⁵ Department of Immunology, School of Basic Medical Science, Anhui Medical University, Hefei, China.

Abstract

Background: Invasion and migration of cancer cells play a key role in lung cancer progression and metastasis. Tumor-associated neutrophils (TANs) are related to poor prognosis in many types of cancer. However, the role of TANs in lung cancer is controversial. In this study, we investigated the effect of TANs on the invasion and migration of lung adenocarcinoma. Methods: Immunohistochemistry was performed to detect the density of infiltrating TANs and the expression of Notch3 in 100 lung adenocarcinoma tissues. Flow cytometry was used to observe the viability of neutrophils, which were isolated from healthy peripheral blood and then exposed to the supernatant of cultured lung adenocarcinoma cell lines. After treating with tumor-associated neutrophils culture supernatant, NeuCS (supernatant of cultured neutrophils), tumor cells culture supernatant, Medium (serum-free medium), respectively, the migration and invasion of the lung cancer cells before and after transfected by si-Notch3 were detected by transwell assay and wound healing assay. Kaplan-Meier plotter (http://kmplot.com/analysis/index.php?p) was used to analyze the prognostic role of the density of TANs on lung adenocarcinoma and TIMER ((http://cistrome.dfci.harvard.edu/TIMER/) was used to detect the expression of Notch3 on lung adenocarcinoma. Results: The infiltration of TANs was observed in the parenchyma and stroma of the lung adenocarcinoma, the density of TANs was positively related to the TNM stage and negatively related to the differentiation and prognosis. Notch3 expression of cancer cells was negatively related to the tumor differentiation and prognosis. Compared to quiescent neutrophils, the viability of TCCS-activated neutrophils was enhanced. Both migration and invasion of A549 and PC9 cells were significantly promoted by TANs, while after knocking down Notch3, the migration and invasion of the cancer cells were not affected by TANs. Bioinformatics analysis showed that the density of TANs and the expression of Notch3 were related to the poor prognosis. Conclusion: The results indicated that lung adenocarcinoma cells promote self-invasion and self-migration by activating neutrophils to upregulate the Notch3 expression of cancer cells. The density of infiltrating TANs may be a novel marker for the poor prognosis of lung adenocarcinoma. Targeting TANs might be a potential therapeutic strategy for lung cancer treatment.

Keywords: Notch3; invasion; lung adenocarcinoma; migration; tumor-associated neutrophils.