人IL-4基因修饰诱导肝母细胞瘤细胞凋亡及分化的研究  

The effects of human IL-4 gene transfer on differentiation and apoptosis in hepatoblastoma cells

作　　者：付劲蓉[1] 李成荣[1] 周玉峰[1] 周雅德[2] 

机构地区：[1]深圳市儿童医院,518026 [2]重庆医科大学附属儿童医院

出　　处：《中华微生物学和免疫学杂志》2003年第2期148-151,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 ( 39470 733)

摘　　要：目的　研究人白细胞介素 4(IL 4)基因修饰对肝母细胞瘤细胞凋亡及分化的影响及可能机制。方法　以逆转录病毒为载体将人IL 4基因导入人肝母细胞瘤细胞系 (HepG2 )细胞。台盼蓝拒染、瑞氏染色、放射免疫测定、流式细胞仪细胞周期分析、原位杂交等方法检测人IL 4基因修饰后细胞形态、甲胎蛋白合成以及原癌基因c fos、c jun、c myc表达的变化。流式细胞仪AnnexinⅤ /PI双染色法及间接免疫荧光染色法检测凋亡细胞及凋亡调控基因p5 3、bcl 2的蛋白表达。结果　 (1)人IL 4基因修饰较空载体修饰及野生型HepG2细胞的细胞周期发生G0 /G1期阻滞 ,甲胎蛋白分泌量及原癌基因c fos、c jun、c myc表达降低 (P <0 .0 0 1)。细胞在形态及功能上趋向正常肝细胞转化 ;(2 )较空载体修饰及野生型HepG2细胞 ,IL 4基因修饰后部分细胞形态上出现核固缩等典型凋亡细胞的特征 ,流式细胞仪亦检测到 18.5 %± 4.7%的凋亡细胞 ;(3)人IL 4基因修饰增加p5 3而抑制bcl 2表达 (P <0 .0 5 )。结论　人IL 4基因修饰可诱导肝母细胞的凋亡及分化 ,诱导凋亡细胞可能与上调p5 3蛋白及抑制bcl 2蛋白表达有关。Objective To investigate the effects of human interleukin-4 (IL-4) gene transfer on differentiation and apoptosis in hepatoblastoma cells line (HepG2) cells. Methods Retroviral vector was employed to introduce human IL-4 gene into hepatoblastoma cell line(HepG2) cells. Trypan blue exclusion, whight dye, radioimmunoassay, in situ hybridization, polymerase chain reaction-enzyme linked immunoadsorbent assay (PCR-ELISA) were used to determine its change in morphology and cell cycle. The apoptosis and the expression of p53, bcl-2 were analyzed by flow cytometry. Results The proliferation of the cells transfered by human IL-4 gene were surpressed (P<0.05) and its shape tended to become relatively mature while the production of α-fetal protein (AFP) expression of protooncogenes such as c-fos, c-jun, c-myc were inhibited (P<0.001); human IL-4 gene transfer could also induce the apoptosis in HepG2 cells, the ratio of apoptosis added up to 18.5%±4.7% (P<0.01) at the same time the expression of p53 and bcl-2 were increased and decreased respectively. Conclusion Human IL-4 gene transfer could induce the apoptosis and differentiation in hepatoblastoma cells, the occurrence of apoptosis was probably related to up-regulated the expression of p53 and down-regulate the expression bcl-2. [

关 键 词：人白细胞介素4 基因修饰 肝母细胞瘤 细胞凋亡 细胞分化 人IL-4 

分 类 号：R735.7[医药卫生—肿瘤]