混合脐血间充质干细胞体外抑制C6胶质瘤细胞增殖和诱导凋亡  

Mechanisms of mixed umbilical cord blood-derived mesenchymal stem cells inhibiting proliferation and inducing apoptosis of C6 glioma cells in vitro

作　　者：焦红亮[1] 王晓宁 杜伟[1] 杨波[1] Jiao Hongliang;Wang Xiaoning;Du Wei;Yang Bo(Department of Neurosurgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Radiology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China)

机构地区：[1]郑州大学第一附属医院神经外科,450052 [2]郑州大学附属郑州中心医院放射科,450007

出　　处：《中华实验外科杂志》2019年第5期873-876,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨脐带血衍生间质干细胞(UCB-MSCs)体外抑制C6胶质瘤细胞增殖、诱导凋亡的机制。方法采用反转录-聚合酶链反应(RT-PCR)法检测C6细胞半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-8、B细胞淋巴瘤/白血病-2相关X蛋白(bax)、B细胞淋巴瘤/白血病-2(bcl-2)和Caspase-3 mRNA水平,膜联蛋白V(Annexin V)-碘化丙锭(PI)双染法检测细胞凋亡。结果混合UCB-MSCs在转录水平上有使Caspase-8、bax、Caspase-3 mRNA表达增高的趋势,bcl-2 mRNA表达降低的趋势。促凋亡蛋白表达量和凋亡细胞数随E/T(E:效应细胞即UCB-MSCs,T:靶细胞即C6细胞)的比例增高而增高,呈现比例依赖关系;E∶T=(5+5)∶1,RT-PCR检测C6细胞Caspase-8、bax、bcl-2和Caspase-3 mRNA相对表达量分别为0.78±0.13、0.28±0.03、0.70±0.08、0.76±0.12。当E/T=0∶1、5∶1、(2.5+2.5)∶1、10∶1、(5+5)∶1时,Annexin V-异硫氰酸荧光素(FITC)和PI双染色检测凋亡C6细胞数量分别为(2.9±0.6)、(3.1±0.5)、(14.5±1.8)、(14.3±2.2)、(22.8±4.5)个。结论经植物凝集素(PHA)刺激后,大剂量UCB-MSCs和混合UCB-MSCs都对体外C6细胞具有抑制增殖作用,而且这种抑制作用可能与诱导凋亡明显相关。Objective To study the mechanisms of the mixed umbilical cord blood-derived mesenchymal stem cells(UCB-MSCs)inhibiting proliferation and inducing apoptosis of C6 glioma cells in vitro.Methods Cysteinyl aspartate-specific protease(Caspase)-8,B cell lymphoma/leukemia-2 associated X protein(bax),B cell lymphoma/leukemia-2(bcl-2)and Caspase-3 protein expression in C6 cells was analyzed by immunohistochemistry and Western blotting,respectively.Caspase-8,bax,bcl-2 and Caspase-3 mRNA levels were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Cell cycle was detected by flow cytometry.Apoptosis of C6 cells was examined by Annexin V-propidium iodide(PI)staining and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)methods,respectively.Results The mixed UCB-MSCs had a tendency to increase the expression of Caspase-8,bax,Caspase-3 mRNA,and decrease the expression of bcl-2 mRNA.The expression of apoptosis protein and the number of apoptosis cells increased with the increase of the ratio of E/T(E:effect cells namely UCB-MSCs;T:target cells namely C6 cells),showing a proportional dependence relationship;E∶T=(5+5)∶1.The relative expression of Caspase-8,bax,bcl-2,and Caspase-3 mRNA in C6 cells was 0.78±0.13,0.28±0.03,0.70±0.08,and 0.76±0.12,respectively.When E/T=0∶1,5∶1,(2.5+2.5)∶1,10∶1,(5+5)∶1,the number of apoptosis C6 cells was(2.9±0.6),(3.1±0.5),(14.5±1.8),(14.3±2.2),and(22.8±4.5)cells,respectively.Conclusion After phytohemagglutinin(PHA)stimulation,the mixed UCB-MSCs could inhibit proliferation of C6 cells in vitro,which was related to induction of apoptosis of glioma cells.

关 键 词：胶质瘤 脐血 间充质干细胞 脱噬作用 增殖 

分 类 号：R739.4[医药卫生—肿瘤]