Effect of Agmatinase on the proliferation, migration and invasion of breast cancer cell
10.3760/cma.j.cn115396-20230912-00059
- VernacularTitle:胍丁胺酶对乳腺癌细胞增殖、迁移及侵袭能力的影响
- Author:
Yongchang GAO
1
;
Chao MA
;
Qingjuan YAO
Author Information
1. 天津医科大学总医院普通外科 天津市普通外科研究所，天津 300052
- Keywords:
Breast neoplasms;
Cell proliferation;
Cell invasion;
Cell migration assays;
Agmatinase;
Biomarkers;
Epithelial-mesenchymal transition
- From:
International Journal of Surgery
2024;51(6):388-393
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of Agmatinase (AGMAT) in breast cancer and its effect on the proliferation, migration and invasion of breast cancer cell lines.Methods:The expression levels of AGMAT in cancer and adjacent tissues of 1 094 breast cancer samples in The Cancer Genome Atlas (TCGA) database were analyzed. And its expression degree was verified in breast cancer cell lines MDA-MB-231, MCF-7, HCC-1937 and T-47D. The expression of AGMAT in breast cancer cells was knocked down by shRNA, and the expression level of AGMAT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the protein expression level was detected by Western blotting assay. Cell count and MTT assay were used to detect cell proliferation. Cell apoptosis and cell cycle changes were detected by flow cytometry. Cell migration ability was detected by cell scratch assay, cell invasion ability was detected by Transwell assay, and the expression level of related proteins in cells was detected by Western blotting assay. Measurement data were expressed as mean ± standard deviation ( ± s), comparisons between multiple groups were performed using ANOVA, pairwise comparisons were performed using independent samples t-test, and Tukey′s post hoc multiple test was used. Results:Analyzing the data of breast cancer samples in TCGA database, it was found that the expression level of AGMAT in cancer tissues was significantly higher than that in adjacent tissues ( FC=10.537, P<0.001). Flow cytometry showed that knocking down the expression of AGMAT inhibited the proliferation of breast cancer cells, and induced cell apoptosis [(3.20±0.10)% vs (6.83±0.06)%, t=62.35, P<0.001], and caused G 1 cell cycle arrest [(49.51±2.22)% vs (31.44±1.67)%, t=42.56, P=0.001]. The results of cell scratch assay and Transwell assay showed that decreased GMAT expression could reduce the cell migration ability [(34.27±1.67) % vs (57.97±0.58) %, t=33.52, P<0.001], invasive ability (163.00±1.77 vs 61.00±0.74, t=52.50, P<0.001). The results of Western blotting assay showed that the protein expression levels of Twist, Vimentin, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) decreased, while the protein expression level of E-cadherin increased. AGMAT was involved in the process of epithelial-mesenchymal transition in breast cancer. Conclusion:AGMAT can be used as a prospective biomarker affecting the invasion, metastasis and therapeutic targets of breast cancer.
