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Expression of mucin 13 in lung adenocarcinoma tissues and its effect on the malignant biological behaviors of A549 cells and the possible mechanism
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Abstract:
Objective: To investigate the expression of mucin 13 (MUC13) in lung adenocarcinoma tissues and its effects on the proliferation, apoptosis, migration, invasion, and epithelial-mesenchymal transition (EMT) of A549 cells as well as the possible mechanism. Methods: The differential expression of MUC13 in lung adenocarcinoma tissues, normal lung tissues, and para-cancerous tissues was analyzed using The Cancer Genome Atlas (TCGA) and high throughput Gene Expression Omnibus (GEO) databases. The mRNA and protein expression levels of MUC13 in human lung adenocarcinoma cells (NCI-H1395, NCI-H1975, H1299 and A549) and normal lung epithelial BEAS-2B cells were detected using qPCR and WB methods. siRNA technology was used to knock down MUC13 expression in A549 cells, and the experimental cells were divided into si-MUC13 group, NC group and si-MUC13+IGF-1 group. The effects of MUC13 knockdown on the proliferation, cell cycle, apoptosis, migration and invasion of A549 cells were detected by clony formation assay, flow cytometry and Transwell assay, respectively. WB assay was used to detect the effect of MUC13 knockdown on the protein expression of E-cadherin, N-cadherin, vimentin, EGFR, p-EGFR, PI3K, p-PI3K, AKT and p-AKT in A549 cells. Results: MUC13 was highly expressed in lung adenocarcinoma tissues and cells at both mRNA and protein levels (all P<0.01),and A549 cells with higher MUC13 expression were selected for subsequent experiments. After knocking down MUC13, the proliferation ability of A549 cells was significantly weakened, the number of cells in G0/G1 phase was significantly increased while the number of cells in G2/M phase and S phase was significantly decreased, the apoptosis rate was significantly increased, and the cell migration and invasion abilities were significantly weakened (all P<0.01); Moreover, the protein expression of E-cadherin in A549 cells was upregulated, while the protein expression of N-cadherin and vimentin was downregulated, and the ratios of p-EGFR/EGFR, p-PI3K/PI3K, and p-AKT/AKT were all reduced (all P<0.01). However, after adding pathway activator IGF-1, the p-EGFR/EGFR, p-PI3K/PI3K, and p-AKT/AKT ratios in A549 cells all increased (all P<0.01). Conclusion: MUC13 is highly expressed in lung adenocarcinoma tissues and cells, and it promotes cell proliferation, migration, invasion and EMT of A549 cells, possibly through the activation of the EGFR/PI3K/AKT signaling pathway.
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