不同人源脐血间充质干细胞抑制胶质瘤细胞增殖及诱导凋亡

Mechanisms of the mixed umbilical cord blood-derived mesenchymal stem cells in inhibiting proliferation and inducing apoptosis of glioma cells

目的探讨不同人源脐血间充质干细胞(UCB-MSCs)体外抑制C6胶质瘤细胞增殖、诱导凋亡的机制。方法采用免疫组织化学法和蛋白质印迹法(Western blot)检测C6细胞B细胞淋巴瘤/白血病-2(bcl-2)和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3蛋白的表达,流式细胞仪检测C6细胞周期。应用SPSS 13.0统计软件分析,实验数据采用均值±标准差(Mean±SD)表示,使用前先行正态性检验和方差齐性检验,符合正态分布并方差齐的,根据数据类型应用参数检验中的单因素方差分析或t检验,不符合正态分布的应用非参数检验中秩和检验。结果不同人源UCB-MSCs对C6细胞在蛋白表达上有使凋亡蛋白Caspase-3表达增高的趋势,抑凋亡蛋白bcl-2表达降低的趋势;不同人源UCB-MSCs诱导C6细胞凋亡,E(效应细胞即UCB-MSCs)/T(靶细胞即EGFP-C6细胞或C6细胞)=0∶1、5∶1、(2.5+2.5)∶1、10∶1、(5+5)∶1作用于的C6细胞,G0/G1期百分比分别为(65.49±1.83)%、(64.83±2.19)%、(70.30±3.21)%、(69.03±2.19)%、(77.63±3.10)%,G0/G1期的C6细胞百分比随着E/T比例增高而增高,呈现比例依赖关系。结论经植物血凝素(PHA)刺激后,大剂量UCB-MSCs和不同人源UCB-MSCs都对体外C6细胞具有抑制增殖作用,而且这种抑制作用与诱导凋亡相关。

Objective To investigate the possible mechanism of mixed umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) in vitro to inhibit proliferation and induce apoptosis of C6 glioma cells. Methods The expression of B cell lymphoma/leukemia-2 (bcl-2) and cysteinyl aspartate-specific protease (Caspase)-3 proteins in C6 cells was detected by immunohistochemistry and Western blotting imprinting method, and C6 cell cycle was detected by flow cytometry. Results Different sources of UCB-MSCs have a tendency to increase the expression of apoptosis protein Caspase-3 in C6 cells and reduce the expression of apoptosis protein bcl-2. Different sources UCB-MSCs induce C6 cell apoptosis, E/T=0∶1, 5∶1,(2.5+ 2.5)∶1, 10∶1,(5+ 5)∶1 C6 cells acting on, The percentages of G0/G1 were (65.49±1.83)%,(64.83±2.19)%,(70.30±3.21)%,(69.03±2.19)%,(77.63±3.10)%, and the percentage of C6 cells in G0/G1 increased with the increase of the E/T ratio, showing a proportional dependence. Conclusion After phytohemagglutinin (PHA) stimulation, large doses of UCB-MSCs and mixed UCB-MSCs have inhibitory effects on proliferation of C6 cells in vitro, and this inhibition may be related to apoptosis induction.