野生型p53基因与双脱水二乙酰卫矛醇联合诱导肝癌HLE细胞凋亡的研究  

Apoptosis of Hepatocarcinoma Cell Line HLE Induced by the Combination of Wild Type P53 Gene and 1,2:5,6-Dianhydro-3,4-diacetylgalactitol

作　　者：赖祥进[1] 曹林枝[1] 周瑛[2] 蓝秀万[1] 廖志红[1] 周素芳[1] 

机构地区：[1]广西医科大学生物化学与分子生物学教研室,广西南宁530021 [2]广西中医药研究所,广西南宁530021

出　　处：《癌症》2004年第10期1139-1143,共5页Chinese Journal of Cancer

基　　金：广西自然科学基金(桂科自No.9912035)~~

摘　　要：背景和目的:p53基因是抑癌基因,研究发现在人类肿瘤中p53基因常发生突变。实验证明野生型p53基因不但抑制肿瘤细胞增殖,而且诱导肿瘤细胞凋亡。但是,单独应用野生型p53基因诱导肿瘤细胞凋亡的作用不很明显。因此,野生型p53基因与药物的联合作用来提高肿瘤细胞凋亡率是目前对肿瘤研究的一新领域。本实验通过转染野生型p53基因联合双脱水二乙酰卫矛醇(1,2:5,6-dianhydro-3,4-diacetylgalactitol,DADAG)作用于肝癌细胞株HLE,研究联合诱导肿瘤细胞的凋亡作用。方法:野生型p53基因通过脂质体Lipofectamine介导转染人肝癌细胞株HLE,同时加入DADAG。96h后,用流式细胞仪、DNA电泳方法进行细胞凋亡分析。结果:流式细胞仪分析细胞凋亡结果:阴性对照组为1.4%、转染pUHD10-3组为3.5%、DADAG处理组为32.6%、转染pUHD10-3-p53组为43.4%、转染p53基因及DADAG处理组为74.6%。DNA电泳发现,转染p53基因及DADAG处理组有到DNA梯形条带。结论:野生型p53基因与DADAG均可诱导人肝癌细胞HLE发生凋亡,转染野生型p53基因与DADAG联合作用,有促进肝癌细胞凋亡的作用。BACKGROUND &OBJECTIVE: P53 gene is a tumor suppressor gene,and its mutation in human tumor cells is frequently observed. Previous studies revealed that wild type p53 (wt-p53) gene can suppress proliferation ,and induce apopto sis of tumor cells. However, the enhancive effect of wt-p53 on apoptosis of tum or cells is not so obvious when it is used alone. Therefore, it is a new field f or tumor research that wt-p53 gene combined with drug is used to enhance apopto sis rate of tumor cells. This study was to investigate the enhancement effect of the combination of wt-p53 and 1,2:5,6-dianhydro-3,4-diacetylgalactitol (DAD AG) on apoptosis of human hepatocarcinoma cell line HLE. METHODS: HLE cells were transfected with pUHD10-3-p53 plasmid contained wt-p53 gene, and treated wit h DADAG. After 96-hour treatment, apoptosis was evaluated by flow cytometry and DNA electrophoresis. RESULTS: The apoptosis rates were: 1.4%in untreated group , 3.5%in pUHD10-3-transfection group, 32.6%in DADAG group, 43.4%in pUHD10-3-p53-transfection group, and 74.6%in pUHD10-3-p53-transfection plus DADAG-treatment group. DNA ladder was observed in pUHD10-3-p53-transfection plus DADAG-treatment group. CONCLUSION: Apoptosis of HLE cells could be induced by b oth wt-p53 gene and DADAG, and the effect was more obvious when HLE cells were treated by the combination of wt-p53 gene and DADAG.

关 键 词：野生型P53基因 转染 HLE 联合诱导 肿瘤细胞凋亡 肝癌 处理 DNA 电泳 发现 

分 类 号：R735.7[医药卫生—肿瘤]