外源野生型p53基因对人胆囊癌GBC-SD细胞生长的作用  

Exogenous wild-type p53 gene suppressing the growth of human gallbladder cancer GBC-SD cell line

作　　者：乌新林[1] 王占民[1] 马道新[2] 杨凤辉[1] 孙建芝[2] 

机构地区：[1]山东大学齐鲁医院普外科 [2]山东大学齐鲁医院血液学研究室,济南250012

出　　处：《中华实验外科杂志》2005年第12期1477-1479,共3页Chinese Journal of Experimental Surgery

基　　金：山东省科技厅发展计划项目(2001BBICJA10)

摘　　要：目的观察外源野生型p53(wtp53)基因对人胆囊癌GBC-SD细胞生长的抑制作用。方法用脂质体介导的转染技术,将含有wtp53的真核表达质粒pCMV-p53导入GBC—SD细胞。用聚合酶链反应(PCR)和逆转录-聚合酶链反应(RT—PCR)证实外源p53基因的整合与表达;用细胞计数和克隆形成实验反映细胞增殖状况;用流式细胞仪检测细胞周期的改变。结果转化细胞系中存在外源p53基因的整合与表达。稳定转染wtp53基因的GBC-SD-wtp53细胞,体外生长速率明显减慢;克隆形成率仅为3．2％,显著低于对照组GBC-SD-mutp53(28％)和亲本GBC-SD细胞 (29％,P<0．01);G0／G1期、S期、G2／M期比例(％)分别为(66．12±4．2、32．87±2．15、1．01± 0．37),与对照组(46．20±5．1、30．78±2．92、23．02±1．87)及亲本细胞(41．25±3．2、40．03±2．09、 18．72±1．05)相比,G0／G1期细胞比例明显增加,G2／M期细胞比例显著减少(P<0．01)。结论外源野生型p53基因的表达能有效抑制人胆囊癌GBC-SD细胞的体外生长。Objective To observe the growth-suppressive effect of exogenous wild-type p53 （wtp53） gene on human gallbladder cancer GBC-SD cell line. Methods Eukaryotic expressing plasmid pCMV-p53 containing human wtp53 gene was introduced by lipofectamine-mediated gene transfection into human gallbladder cancer GBC-SD cell line. The presence apd expression of exogenous p53 gene was confirmed by PCR and RT-PCR, the ability of cell proliferation was assessed using the cell counting and the colony form assay;the cell cycle distribution was detected by flow cytometry. Results The introduction and expression of exogenous p53 gene in transfected cell lines was proved. The cell growth rate and the ability to form colony were remarkably decreased in GBC-SD-wtp53. Cloning efficiency for GBC-SDwtp53 was 3.2 %, which was obviously lower than that for control GBC-SD-mutp53 group （28 % ） and GBC-SD group （29%） （P〈0.01）. The percent of G0/G1, S and G2/M period was （66.12 ± 4.2） %, （32.87 ± 2.15） %, （1.01 ± 0.37） % respectively. Compared with GBC-SD-mutp53 cell and GBC-SD cell, the percent of G0/G1 period was significantly increased and the percent of G2/M period was obviously decreased （P 〈 0.01 ）. Conclusion The expression of exogenous wtp53 gene could effectively inhibit the growth of human gallbladder cancer GBC-SD cell lines in vitro.

关 键 词：p53 胆囊肿瘤 转染 基因治疗 细胞生长 

分 类 号：R735.8[医药卫生—肿瘤]