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Lung cancer has been the leading cause of cancer incidence and mortality in China for years. Benzo[ a ]pyrene (BaP) is a well-known carcinogen for lung cancer. To understand alternation of key proteins and their role in BaP-induced lung cancer, we compared proteome profiles between BaP-transformed 16HBE cell line T-16HBE-C1 (THBEc1) cells and control using label-free quantitative proteomic analysis. Forkhead box protein A1 (FOXA1) was selected and evaluated for its potential role in BaP-induced carcinogenesis in vitro and in vivo. Relationship between FOXA1 expression and survival of lung cancer patients were examined via The Cancer Genome Atlas (TCGA) database. A total of 183 differentially expressed proteins were identified, with 67 proteins including FOXA1 up-regulated and 116 proteins down-regulated in THBEc1 cells. Differentially expressed proteins mainly functioned in basic cellular metabolism, tumor related pathways and regulation of transcription factors. FOXA1 knockout inhibited colony formation and migration of THBEc1 cells in vitro. FOXA1 knockout inhibited tumor growth and metastasis in BALB/c-nude mice after subcutaneous and tail vein injection of FOXA1 knockout THBEc1 cells, respectively. FOXA1 mRNA expression was higher in tumor tissues for patients with lung squamous cell carcinoma, but not associated with survival of lung cancer patients. Our findings revealed oncogenic role of FOXA1 in BaP-induced lung cancer and improved understanding of mechanism in BaP-induced carcinogenesis. • There are significant protein expression changes in BaP-transformed 16HBE cells. • FOXA1 knockout inhibited colony formation and migration of THBEc1 cells. • FOXA1 knockout inhibited tumor growth and metastasis of THBEc1 cells. • FOXA1 is a key protein in BaP-induced lung cancer. [ABSTRACT FROM AUTHOR]

• The differentially expressed miRNAs between THBEc1 and 16HBE cells might participate in BaP-induced carcinogenesis. • MiR-152-3p/ ALCAM , miR-142-5p/ ALCAM and miR-211-5p/ ALCAM axes may be involved in BaP-induced carcinogenesis. • ALCAM plays the role of promoting tumor growth and inhibiting metastasis. Benzo[a]pyrene (BaP) is a potent carcinogen and microRNAs (miRNAs) may play an important role in carcinogenesis. Activated leukocyte cell adhesion molecule (ALCAM) was up-regulated in BaP-transformed 16HBE cell line (THBEc1), and may be a key molecule for THBEc1 cells to gain and maintain the malignant phenotype. Here we screened the differentially expressed miRNAs which resulted in up-regulation of ALCAM in THBEc1 cells by comparing miRNA expression profiles between THBEc1 and 16HBE (HBE) cells. Results showed that a total of 555 miRNAs differentially expressed between THBEc1 and HBE cells, of which 351 miRNAs were down-regulated and 204 miRNAs were up-regulated in THBEc1 cells. MiR-152-3p, miR-142-5p and miR-211-5p down-regulated in THBEc1 cells were demonstrated to participate in the regulation of ALCAM. With THBEc1 as a tumor cell model, we determined the role of ALCAM in tumor growth and metastasis employing two ALCAM knockout THBEc1 cell lines via CRISPR/Cas9 technology. Results showed that ALCAM knockout inhibited colony formation and tumor growth, but enhanced cell migration and lung metastasis of THBEc1 cells. In conclusion, miR-152-3p/ ALCAM , miR-142-5p/ ALCAM and miR-211-5p/ ALCAM axes may be involved in BaP-induced carcinogenesis. BaP might induce up-regulation of ALCAM via inhibiting miR-152-3p, miR-142-5p and miR-211-5p, which in turn allows ALCAM to exert its role promoting cell proliferation and tumor growth, and suppressing cell migration and metastasis. [ABSTRACT FROM AUTHOR]

Benzo[ a ]pyrene (BaP) is a strong carcinogen for lung cancer, and forkhead-box A1 (FOXA1) plays an oncogenic role in BaP-transformed cell THBEc1. To explore the remodeling of metabolic pattern caused by BaP-induced transformation and the possible role FOXA1 might play in it, we compared metabolic patterns between THBEc1 cells and control using untargeted metabolomics and lipidomics analysis, and determined the effects of FOXA1 knockout on the metabolic pattern of THBEc1 cells. Metabolomics and lipidomics identified a total of 15 and 46 differential metabolites and lipids between THBEc1 and 16HBE cells, respectively, and a total of 4 and 1 differential metabolites and lipids between FOXA1 knockout cell THBEc1-Δ FOXA1 -c34 and control cell THBEc1-ctrl, respectively. Analysis results of metabolites and metabolic pathways indicated the metabolic pattern remodeling may be related to the alteration in glucose metabolism during BaP-induced transformation. Western blotting revealed the up-regulation of enolase-2 (ENO2), pyruvate carboxylase (PCB), aconitase-2 (ACO2) and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) (Thr202/Tyr204), the down-regulation of succinate dehydrogenase complex subunit A (SDHA) and phosphoenolpyruvate carboxykinase 2 (PCK2) in THBEc1 cells. The detection results of metabolites related to glucose metabolism demonstrated the decreasing of lactic acid content in cells, lactic acid production in culture medium and citric acid content in mitochondria, and the increasing of ATP production of THBEc1 cells. FOXA1 knockout partially reversed the changes of ENO2, SDHA, PCK2 and p-ERK1/2 (Thr202/Tyr204) levels, lactic acid release, citric acid content in mitochondria of THBEc1 cells. In conclusion, FOXA1 knockout partially reversed the remodeling of glucose metabolism caused by BaP-induced malignant transformation. Our findings provide a clue for the possible role of FOXA1 in glucose metabolism regulation. [Display omitted] • There are significant metabolic pattern changes in BaP-transformed cell THBEc1. • Metabolic pattern remodeling of THBEc1 cells is related to glucose metabolism. • FOXA1 knockout partially reversed the remodeling of glucose metabolism in THBEc1 cells. • MAPK signaling pathway is activated in THBEc1 cells. • FOXA1 knockout decreased the levels of p-ERK1/2 (Thr202/Tyr204) in THBEc1 cells. [ABSTRACT FROM AUTHOR]