自杀基因修饰内皮祖细胞对肝细胞癌体外作用的效应

目的：研究自杀基因胞嘧啶脱氨酶（CD）基因转染的小鼠骨髓源性内皮祖细胞（EPCs）联合酶前体药物5-氟胞嘧啶（5-FC）对小鼠肝癌细胞株H22的体外抗增殖作用。方法：分离普通小鼠骨髓源性EPCs，培养鉴定，Polybrene技术转染含CD基因的慢病毒载体重组体plenti6．3-EGFP-CD至EPCs，Western blotting检测目的蛋白的表达。利用Transwell小室共培养体系，用转染CD基因的EPCs处理H22细胞。 CCK-8法检测H22细胞增殖变化，流式细胞仪检测细胞凋亡水平。结果：成功转染CD基因至EPCs；CCK-8法检测显示随着5-FC浓度增加，细胞增殖逐渐下降，5-FC浓度达100 mg· L-1时，肿瘤细胞增殖抑制率达到（54．74±5．38）％（P＜0．05），细胞平均凋亡率为（48．71±4．62）％（P＜0．05）。结论：CD基因修饰的EPCs 联合5-FC体外可明显抑制肝癌H22细胞增殖，诱导肿瘤细胞凋亡。

Effects of endothelial progenitor cells modified by suicide gene on proliferation of mice hepatoma cells in vitro

Objective：To investigate the suppression of the proliferation of hepatoma H 22 cells treated through mice marrow-derived endothelial progenitor cells（EPCs） modified by cytosine deaminase （CD） gene and enzyme prodrug 5-fluorocytosine （5-FC） in vitro.Methods： EPCs were isolated from mice bone marrow .After cultured and identified, EPCs were transfected by lentiviral vector carrying CD gene （plenti6.3-EGFP-CD ） using Polybrene technique .Expression of target protein was tested by Western blotting .H22 cells were treated by the EPCs carrying CD gene using Transwell cells co-culture system .CCK-8 method was used to detect the proliferation of H 22 cell. The tumor apoptosis was tested by flow cytometry .Results： The transfection of CD gene to EPCs was detected successfully .CCK-8 assay showed that cell proliferation gradually decreased along with increasing of 5-FC concentration .When 5-FC reached 100 mg· L-1 , the proliferation inhibition of tumor cells was about （ 54 .74 ± 5.38）%（P〈0.05） and the average apoptosis rate was （48.71 ±4.62）%（P 〈0.05）.Conclusion： EPCs modified by CD gene with 5-FC can effectively inhibit hepatoma H 22 cells proliferation and induce cells apoptosis in vitro.