Effects of mesenchymal stem cell exosomes on biological behavior of esophageal carcinoma ECA109 cells

doi:10.11958/20241497

Abstract

Abstract:

Objective To explore the effects of exosomes of mesenchymal stem cells (MSC-Exos) on the proliferation, apoptosis, migration and invasion of esophageal cancer ECA109 cells. Methods Human umbilical cord mesenchymal stem cells were cultured, exosomes were extracted and isolated, and identified by transmission electron microscopy. The nanoparticle size determination and protein characterization (TSG101, CD63) were measured by transmission electron microscope. There were the MSC-Exo group (MSC-Exos co-cultured with esophageal cancer ECA109 cells) and the blank control group (only esophageal cancer ECA109 cells), and cells were cultured for 0, 24 and 48 h, respectively. CCK-8 proliferation test and scratch test were used to detect the proliferation and migration ability of esophageal cancer EAC109 cells in each group, respectively. After 48 h of culture, cell apoptosis and cell cycle changes were detected by flow cytometry. The protein expression levels of phosphoinositol 3-kinase phosphorylation (p-PI3K), rabbit phosphorylated protein kinase B phosphorylation (p-Akt) and β-catenin were detected by Western blot assay. Results After identification, the obtained MSC-Exos meeted the required standard. Transmission electron microscopy, particle size measurement and marker protein results confirmed that the extracted exosomes of mesenchymal stem cells meeted the identification criteria. At 0 h of cell culture, there were no significant differences in cell proliferation and migration healing rate between the two groups (P>0.05). After 24 h culture, the cell proliferation ability was lower in the MSC-Exo group than that of the blank control group (P<0.05). After 48 h culture, the cell proliferation and migration healing rate were lower in the MSC-Exo group than those of the blank control group (P<0.05). The apoptosis rate of the MSC-Exo group was higher than that of the blank control group, and the proportion of G2+S phase cells was lower than that of blank control group (P<0.05). The expression levels of p-PI3K, p-Akt and β-catenin protein were significantly lower in the MSC-Exo group than those in the blank control group (P<0.05). Conclusion MSC-Exos can inhibit the proliferation and migration of esophageal cancer cells and promote cell apoptosis. The inhibitory effect of MSC-Exos on esophageal cancer cells may be related to inhibiting the activation of PI3K and Akt protein and the down-regulating expression of β-catenin protein.

Key words: mesenchymal stem cells, exosomes, esophageal neoplasms, cell proliferation, cell movement, apoptosis

CLC Number:

R735.1

Figures/Tables 8

Fig.1 Identification of exomes

Fig.2 Effect of MSC-Exo on cell migration of esophageal carcinoma ECA109

Tab.1 Comparison of proliferation and scratch migration of ECA109 cells between different time points of two groups

组别	细胞增殖能力（OD₄₅₀）
组别	0 h	24 h	48 h
空白对照组	0.52±0.01	0.89±0.08	2.04±0.16
MSC-Exo组	0.54±0.02	0.65±0.03	1.14±0.15
t	1.543	6.165^＊	9.080^＊
组别	迁移愈合率/%
组别	0 h	24 h	48 h
空白对照组	0.77±0.25	1.29±0.33	2.97±0.38
MSC-Exo组	0.62±0.26	1.22±0.41	1.62±0.62
t	1.212	1.276	2.376^＊

Tab.2 Comparison of apoptosis and cell cycle of ECA109 cells between the two groups

组别	细胞凋亡率（Q2+Q4期）	G2+S期细胞比例
空白对照组	7.23±0.86	55.17±9.60
MSC-Exo组	10.66±0.37	39.26±0.80
t	6.315^＊	2.859^＊

Fig.3 Effects of MSC-Exo on apoptosis of ECA109 cells

Fig.4 Effects of MSC-Exo on cell cycle of ECA109 cells

Tab.3 Comparison of expression levels of p-PI3K, p-Akt and β-catenin between the two groups of esophageal cancer cells

组别	p-PI3K	p-Akt	β-catenin
空白对照组	1.29±0.03	0.61±0.02	1.07±0.02
MSC-Exo组	0.90±0.01	0.51±0.01	0.68±0.01
t	20.003^＊	8.091^＊	23.839^＊

Fig.5 Western blot results of expression levels of PI3K, Akt and β-catenin protein in esophageal cancer cell ECA109

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