脂质体介导的c—myc反义硫代寡核苷酸联合5-氟尿嘧啶诱导肝癌细胞HEPG-2的凋亡  

Introduction of apoptosis of human hepatoma cells HEPG-2 with liposome-mediated c-myc antisense phosphorothioate oligodeoxynucleotide and 5-FU

作　　者：郭天康[1] 蔡辉[1] 孙延庆[1] 李振军[1] 魏丽[1] 

机构地区：[1]甘肃省人民医院普外科,兰州 730000

出　　处：《中华普通外科杂志》2011年第3期246-250,共5页Chinese Journal of General Surgery

基　　金：基金项目：甘肃省科学事业费研究基金资助项目（Qs031-c33-02）

摘　　要：目的观察c-myc基因反义硫代寡核苷酸（antisense oligodeoxynucleotide，ASODN）和5-氟尿嘧啶（5-fluorouracil，5-FU）对HEPG-2肝癌细胞增殖及c-myc基因表达的抑制作用，探讨c-mycASODN作为药物进行肝癌基因与药物综合治疗的可行性。方法应用脂质体介导的c—myc反义硫代寡核苷酸和5-FU联合作用于HEPG-2肝癌细胞，M1Tr法检测细胞增殖状态，RT—PCR和免疫组织化学方法检测c—myc堆因表达抑制情况，流式细胞仪进行细胞周期分析，细胞荧光染色及基因组电泳观察细胞凋亡情况。结果e．mycASODN转染HEPG-2肝癌细胞后，细胞生长受抑制，其抑制作用有时问性及剂量依赖性（P=0．02，P=0．01），增殖能力减弱，同时ASODN能在转录和翻译水平上抑制c—myc基因的表达；5．Fu在10μmol／L时可明显诱导肝癌细胞凋亡；将c—mycASODN与5-Fu联合应用，则表现出对细胞凋亡的协同诱导作用（P=0．01）。结论脂质体介导的c—mycASODN及5-yulec—myc基因的表达具有明显的抑制作用，能抑制肝癌细胞增殖。c—mycASODN能够增加肝癌细胞对5－FU的敏感性。Objective To study the inhibition effect of c-myc ASODN （ antisense oligodeoxynucleotide） and 5-FU （5-fluorouracil） on the expression of c-mye gene and the proliferation of human hepatoma ceils HEPG-2. Methods After treated by liposome mediated c-myc antisense phosphorothioate oligodeoxynucleotide （APSODN） and 5-FU, the growth inhibition rate was detected by M＇I~ assay, the expression of c-myc mRNA was detected by RT-PCR and immunohistocehemical methods HEPG-2 cells. The cell cycle was analyzed by flow cytometric analysis. The morphological changes were observed by fluorescence staining and cellular genome electrophoresis. Results After sealing c-myc gene with ASODN, the growth of cells was repressed and the effect was time-dependent and dose-dependent （ P = 0.02 ）. The ability of proliferation decreased, the expression of c-myc gene was inhibited on transcription and translation levels ; 5-FU can induce apoptosis of hepatoma cells HEPG-2 dramatically with the dose of 10 μmo]/L, when treated by both c-myc ASODN and 5-FU, HEPG-2 cells was induced apoptosis in a cooperative style （ P = 0. O1 ）. Conclusions The liposome mediated c-myc （APSODN） and 5-FU can inhibit the proliferation of HEPG-2 cells by inhibiting the expression of c-myc gene and can induce apoptosis of hepatoma cells HEPG- 2 in a cooperative style, c-myc （APSODN） can increase the sensitivity of 5-FU to hepatoma cells and decrease the effective concentration of 5-FU.

关 键 词：癌 肝细胞 细胞凋亡 基因 c—myc 

分 类 号：R735.7[医药卫生—肿瘤]