自杀基因修饰内皮祖细胞对肝细胞癌体外作用的效应-东南大学学报医学版

自杀基因修饰内皮祖细胞对肝细胞癌体外作用的效应

单位：1. 东南大学附属中大医院肿瘤中心, 江苏南京 210009;
2. 东南大学江苏省分子影像与功能影像重点实验室, 江苏南京 210009

分类号：R73-3

出版年·卷·期（页码）：2014·33·第三期（291-296）

摘要：

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。

Objective:To investigate the suppression of the proliferation of hepatoma H22 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 H22 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 H22 cells proliferation and induce cells apoptosis in vitro.

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