野生型p53基因对不同内源性p53状态的肝癌细胞株生长的影响  

Therapeutic potential of recombinant adenovirus expressing p53 on hepatocellular carcinoma cells possessing different p53 functional status

作　　者：郭英[1] 王坤[2] 陈积圣[2] 

机构地区：[1]中山医科大学第三附属医院外科,广州510630 [2]中山医科大学第二附属医院肝胆外科,广州510630

出　　处：《中华普通外科杂志》2001年第12期722-724,共3页Chinese Journal of General Surgery

基　　金：国家自然科学基金 ( 396 70 70 5 )资助

摘　　要：目的探讨外源性野生型p5 3(wild typep5 3,WT p5 3)基因对具有不同内源性p5 3功能状态肝癌细胞生长的影响。方法通过表达WT p5 3的重组腺病毒载体 (Ad p5 3) ,将p5 3基因导入具有不同内源性p5 3功能状态的肝癌细胞株Bel 740 2、QGY 770 1和PLC/PRF/ 5中 ,用MTT法检测Ad p5 3对各细胞株生长的影响 ,流式细胞仪分析各组细胞中DNA含量及凋亡的比例。结果当取 40、2 0 0、80 0MOI值的表达p5 3的腺病毒分别转染PLC/PRF/ 5、Bel 740 2和QGY 770 1细胞时 ,72h后行MTT检测 ,OD值分别为 :在PLC/PRF/ 5细胞 ,空载体 (Ad LacZ)组 0 98± 0 16 ,Ad p5 3组 0 0 2± 0 0 0 (P <0 0 1) ;在Bel 740 2细胞 :Ad LacZ组 1 0 4± 0 2 3,Ad p5 3组 0 10± 0 0 2 (P <0 0 1) ;而在QGY 770 1细胞 :Ad LacZ组 1 88± 0 2 4,Ad p5 3组 1 86± 0 19(P >0 0 5 )。细胞周期分析显示 ,在PLC/PRF/ 5 ,表现为既诱导细胞凋亡 ,又诱导细胞周期阻滞。在Bel 740 2 ,表现为诱导细胞周期阻滞。结论外源性p5 3对肝癌细胞生长的抑制可能与内源性p5 3功能状态无关。Objective To determine the effect of exogenous wild-type p53(WT-p53) on the growth of hepatocellular carcinoma (HCC) cell lines possessing different p53 functional status.Methods With recombinant adenoviral vector expressing WT-p53(Ad-p53),p53 gene was transfected into HCC cell lines:Bel-7402,QGY-7701, and PLC/PRF/5.The cytotoxicities of Ad-p53 to these cell lines were measured by MTT assay,cell growth properties and cell cycle pattern were assessed by flow cytometry.Results The OD of PLC/PRF/5, Bel-7402,and QGY-7701 cells transfected with Ad-p53 of 40,200,and 800 MOI were 0.02±0.00 vs 0.98±0.16(P<0.01), 0.10±0.02 vs 1.04±0.23(P<0.01), and 1.86±0.19 vs 1.88±0.24(P>0.05),compared with control groups respectively. Analysis of flow cytometry showed higher G2/M? lower S and apoptosis ratio in PLC/PRF/5 cells,while higher G0/G1 ratio and lower S ratio in Bel-7402 cells. ConclusionReplication-deficient adenoviral vector expressing WT-p53 may be useful for gene therapy in HCC regardless the status of endogenous p53.

关 键 词：肝细胞 基因 P53 基因表达调控 腺病毒科 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]