间充质干细胞外泌体对食管癌ECA109细胞生物学行为的影响

doi:10.11958/20241497

天津医药 ›› 2025, Vol. 53 ›› Issue (2): 113-117.doi: 10.11958/20241497

• 细胞与分子生物学 • 下一篇

间充质干细胞外泌体对食管癌ECA109细胞生物学行为的影响

马莉莉¹(), 李子沐², 王亮¹, 许彭³, 李秀梅³^,^△()

1 新疆医科大学第五附属医院检验科（邮编830011）
2 成都大学临床医学2020级
3 新疆医科大学基础医学院

收稿日期:2024-10-14 修回日期:2024-12-04 出版日期:2025-02-15 发布日期:2025-02-26
通讯作者: ^△ E-mail：641251840@qq.com
作者简介:马莉莉（1984），女，副主任技师，主要从事肿瘤发病分子机制方面研究。E-mail：venuslilyma@163.com
基金资助:
新疆维吾尔自治区自然科学基金项目(2021D01C290)

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

MA Lili¹(), LI Zimu², WANG Liang¹, XU Peng³, LI Xiumei³^,^△()

1 Department of Clinical Laboratory, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China
2 Grade 2020 of Clinical Medicine, Chengdu University
3 the School of Basic Medicine of Xinjiang Medical University

Received:2024-10-14 Revised:2024-12-04 Published:2025-02-15 Online:2025-02-26
Contact: ^△ E-mail: 641251840@qq.com

马莉莉, 李子沐, 王亮, 许彭, 李秀梅. 间充质干细胞外泌体对食管癌ECA109细胞生物学行为的影响[J]. 天津医药, 2025, 53(2): 113-117.

MA Lili, LI Zimu, WANG Liang, XU Peng, LI Xiumei. Effects of mesenchymal stem cell exosomes on biological behavior of esophageal carcinoma ECA109 cells[J]. Tianjin Medical Journal, 2025, 53(2): 113-117.

摘要/Abstract

摘要：

目的探讨间充质干细胞外泌体（MSC-Exos）对食管癌ECA109细胞增殖、凋亡及迁移能力的影响。方法培养人脐带间充质干细胞，提取并分离外泌体，采用透射电镜、纳米粒径测定及表征蛋白（TSG101、CD63）进行鉴定。实验设MSC-Exo组（MSC-Exos与食管癌ECA109细胞共培养）和空白对照组（仅培养食管癌ECA109细胞），分别培养0、24、48 h，采用CCK-8增殖、划痕实验分别检测各组食管癌EAC109细胞增殖、迁移能力；培养48 h后，采用流式细胞仪检测细胞凋亡及细胞周期变化，Western blot检测磷酸化磷酸肌醇3-激酶（p-PI3K）、兔源磷酸化蛋白激酶B（p-Akt）、β-catenin蛋白的表达。结果经鉴定，所得MSC-Exos符合要求标准。细胞培养0 h时，2组细胞增殖能力和迁移愈合率差异均无统计学意义（P>0.05）；培养24 h时，MSC-Exo组细胞增殖能力低于空白对照组（P<0.05）；培养48 h时，MSC-Exo组细胞增殖能力和迁移愈合率均低于空白对照组（P<0.05）。MSC-Exo组细胞凋亡率高于空白对照组，G2+S期细胞比例低于空白对照组（P<0.05）。MSC-Exo组p-PI3K、p-Akt和β-catenin蛋白表达水平均低于空白对照组（P<0.05）。结论 MSC-Exos能够抑制食管癌细胞增殖、迁移，促进细胞凋亡，其对食管癌细胞的抑癌作用可能与抑制PI3K、Akt蛋白的活化、下调β-catenin蛋白表达有关。

关键词: 间质干细胞, 外泌体, 食管肿瘤, 细胞增殖, 细胞运动, 细胞凋亡

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

中图分类号:

R735.1

图/表 8

图1 外泌体鉴定 A：外泌体的透射电子显微镜图（×30 000）；B：纳米颗粒粒径分析跟踪；C：Western blot检测外泌体标志蛋白的表达。

Fig.1 Identification of exomes

图2 MSC-Exo对食管癌ECA109细胞迁移的影响

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

表1 2组在培养不同时点的细胞增能力和迁移愈合率比较（n=3，$\bar{x}±s$）

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^＊

表2 2组细胞凋亡和细胞周期的比较（n=3，%，$\bar{x}±s$）

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^＊

图3 MSC-Exo对食管癌细胞凋亡的影响

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

图4 MSC-Exo对食管癌细胞周期的影响

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

表3 2组食管癌细胞p-PI3K、p-Akt、β-catenin 蛋白表达水平比较（n=3，$\bar{x}±s$）

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^＊

图5 Western blot检测食管癌细胞PI3K、Akt及β-catenin蛋白的表达

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

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间充质干细胞外泌体对食管癌ECA109细胞生物学行为的影响

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

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