Major tobacco carcinogen benzo(a)pyrene induces infiltration of M2 type macrophages by upregulating cyclooxygenase-2 expression in esophageal squamous carcinoma

doi:10.3969/j.issn.1006-7795.2023.05.008

Abstract

Abstract: Objective To investigate the potential mechanism of increased infiltration of M2 tumor-associated macrophages in the tumor microenvironment of esophageal squamous carcinoma patients with a history of smoking. Methods Differential expression of M2 tumor-associated macrophages in esophageal squamous carcinoma patients with/without a history of smoking was examined by immunohistochemistry and CIBERSORT database analysis. The expression of cyclooxygenase-2（COX2）involved in the regulation of macrophage recruitment and polarization was detected in tumor tissues of esophageal squamous cell carcinoma（ESCC）patients with a history of smoking by The Cancer Genome Atlas (TCGA)database. Esophageal squamous carcinoma cell lines were treated with benzo(a)pyrene, the major carcinogen in tobacco. Changes in the expression of COX2 of ESCC cell lines were detected by Western blotting and real time quantitative PCR (RT-qPCR). The effect on macrophage recruitment by tumor cell conditional medium was assessed by Transwell assay before and after benzo(a)pyrene stimulation. A co-culture system of tumor cell conditional medium and macrophage was established. Western blotting and RT-qPCR were used to evaluate the expression changes of M2 tumor associated macrophages （TAM）markers. Macrophage morphological changes were observed to assist in determining their polarization. Results The infiltration of M2 macrophages was significantly increased in patients with a history of smoking. COX2 was significantly increased in esophageal squamous carcinoma cells stimulated by benzo(a)pyrene, the major carcinogen in tobacco. The recruitment of macrophages by tumor cell conditional medium was significantly increased after stimulation with benzo(a)pyrene. The treatment of macrophages with tumor cell conditional medium after stimulation with benzo(a)pyrene induced their polarization towards M2 macrophages. Conclusions Stimulation of esophageal squamous carcinoma cell lines with benzo(a)pyrene induces macrophages to enter the tumor microenvironment and induces polarization towards M2 tumor-associated macrophages.

Key words: esophageal squamous cell carcinoma, smoking, benzo(a)pyrene, cyclooxygenase-2, tumor associated macrophages

CLC Number:

R735.1

Xiao Zeru, Yan Rui, Liang Ziwei, Ge Yang, An Guangyu. Major tobacco carcinogen benzo(a)pyrene induces infiltration of M2 type macrophages by upregulating cyclooxygenase-2 expression in esophageal squamous carcinoma[J]. Journal of Capital Medical University, 2023, 44(5): 753-761.

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