血小板对胃癌间质干细胞生物学特性的影响

Effect of platelets on the biological characteristics of gastric cancer mesenchymal stem cells

目的观察血小板对胃癌间质干细胞(gastric cancer mesenchymal stem cells,GC-MSCs)的生物学特性以及对GC-MSCs促进肿瘤细胞迁移能力的影响。方法从健康志愿者静脉血标本中分离血小板;体外分离培养人GC-MSCs并分为对照组和GC-MSCs+血小板组,用流式细胞术检测GC-MSCs的细胞表型;成脂、成骨实验检测GC-MSCs的成脂、成骨能力;细胞计数法和Transwell实验检测GC-MSCs的增殖和迁移能力;将GC-MSCs与血小板或肿瘤细胞上清处理后的血小板共培养,western blot检测α-SMA蛋白的表达水平;将SGC-7901与GC-MSCs的细胞培养上清和血小板处理后的GC-MSCs上清共培养,Transwell实验检测SGC-7901的迁移能力。结果 GC-MSCs+血小板组的成脂、成骨能力增强,且增殖能力(24 h,t=3.88,P<0.05; 48 h,t=5.93,P<0.05; 72 h,t=4.35,P<0.05)和迁移能力(t=4.03,P<0.05)也明显加强; SGC-7901上清处理血小板组与单独血小板组的α-SMA蛋白表达水平高于对照组; SGC-7901与GC-MSCs上清或血小板处理后的GC-MSCs上清共培养后,其迁移能力增强(F=42.92,P<0.05)。结论血小板可促进GC-MSCs增殖、迁移及成脂、成骨分化能力,并且可增强GC-MSCs促进肿瘤细胞迁移的能力。

Objective To investigate the effects of platelets on the biological characteristics of gastric cancer mesenchymal stem cells (GC-MSCs) and the ability of GC-MSCs to promote the migration of tumor cells. Methods Platelets were isolated from venous blood samples of healthy volunteers. Human GC-MSCs were isolated and cultured in vitro and divided into the control group and GC-MSCs + platelets group. The phenotype of GC-MSCs was detected by flow cytometry. The adipogenic and osteogenic abilities of GC-MSCs were detected by the adipogenic and osteogenic experiments. The proliferation and migration abilities of GC-MSCs were determined by the cell counting and Transwell assay. After GC-MSCs were co-cultured with platelets or platelets treated with the conditioned medium of SGC-7901 cells,the expression level of α-SMA in GC-MSCs was detected by western blot. The gastric cancer SGC-7901 cells were cocultured with the conditioned medium of GC-MSCs or platelet-treated GC-MSCs,and then the migration ability of SGC-7901 cells were detected by the Transwell assay. Results Compared with the control group,the adipogenic and osteogenic abilities and the proliferation (24h,t= 3.88,P<0.05; 48 h,t= 5.93,P<0.05; 72 h,t= 4.35,P<0.05) and migration abilities (t= 4.03,P<0.05) of GC-MSCs in GC-MSCs + platelets group were enhanced obviously. The expression levels of α-SMA in GC-MSCs treated with platelets or platelets treated with the conditioned medium of SGC-7901-CM were higher than that in the control group. After SGC-7901 cells were co-cultured with the conditioned medium of GC-MSCs or platelet-treated GC-MSCs,their migration abilities were enhanced obviously (F=42.92, P<0.05) . Conclusion Platelets may promote the proliferation,migration,adipogenic and osteogenic differentiation of GC-MSCs,and enhance the ability of GC-MSCs to promote the migration of tumor cells.