MFAP5 inhibits the malignant progression of endometrial cancer cells in vitro

Abstract

To investigate the biological role of MFAP5 in endometrial cancer (EC). HEC-1-A and Ishikawa cells overexpressing MFAP5 were created. Cell proliferation, apoptosis, migration, and invasion were evaluated using CCK8, colony formation, flow cytometry, and transwell assays. A western blot was used to analyze the expression of markers affiliated with the epithelial-mesenchymal transition process and AKT/mTOR pathway. As a result, MFAP5 was found to be down-regulated in EC. Overexpression of MFAP5 suppressed proliferation and promoted apoptosis of HEC-1-A and Ishikawa cells, as evidenced by the inhibition of cell viability and colony formation, and the increase in cell apoptosis rate. Besides, overexpression of MFAP5 attenuated the abilities of cell migration and invasion, as well as reduced MMP2 and MMP9 protein expression. Furthermore, E-cadherin protein level was elevated, while N-cadherin and α-SMA protein levels were decreased, and the phosphorylation of AKT and mTOR was reduced in cells overexpressing MFAP5. Our findings indicate that MFAP5 overexpression inhibits the malignant behaviors of EC cells, possibly by blocking the AKT/mTOR pathway, suggesting that MFAP5 may be a new therapeutic target for EC.

Keywords: AKT/mTOR; EMT; MFAP5; endometrial cancer; proliferation.

Figure 1

MFAP5 is down-regulated in EC. (a) Analysis of MFAP5 expression in 174 UCEC tumors on TCGA. (b) MFAP5 protein expression in cells was analyzed with the western blot method. ^* p < 0.05, ^** p < 0.01, and ^*** p < 0.001.

Figure 2

MFAP5 overexpression inhibits proliferation and promotes apoptosis of EC cells. (a) Overexpression efficiency of MFAP5 was detected by western blot. (b) Cell viability was evaluated using the CCK8 assay. (c) Cell clonogenic ability was examined by colony formation assay. (d) Apoptosis of cells was analyzed by flow cytometry. ^*** p < 0.001.

Figure 3

MFAP5 overexpression attenuates migration and invasion of EC cells. (a) Cell migration and invasion were assessed by transwell assays. (b) Western blot was used to detect MMP2 and MMP9 protein expression. ^** p < 0.01 and ^*** p < 0.001.

Figure 4

MFAP5 overexpression suppresses EMT in EC cells. E-cadherin, N-cadherin, and α-SMA protein expression in cells was tested through western blot assays. ^*** p < 0.001.

Figure 5

MFAP5 overexpression inactivates the AKT/mTOR pathway. Protein expressions of AKT, p-AKT, mTOR, and p-mTOR were determined by western blot assays. ^* p < 0.05 and ^*** p < 0.001.

Figure 6

Schematic diagram of MFAP5 in EC via AKT/mTOR pathway. MFAP5 overexpression inhibited the activation of the AKT/mTOR pathway, thereby suppressing the proliferation, migration, invasion, and EMT of EC cells.