摘要
目的 研究阳离子脂质体Lipofectin(Lip)介导的DNA 聚合酶α(DNA- polα) 反义寡核苷酸(ASODN)体外抑制人胃癌细胞的生长。方法 合成与人pol α催化亚基mRNA 翻译起始区18 个碱基互补的寡核苷酸(ODN) ,与Lip 混合制备Lip+ ODN复合物,作用于MGC-803、SGC-7901 人胃癌细胞24 h 后,细胞换液,继续培养24 h,用噻唑蓝(MTT)比色法测细胞存活状态,流式细胞术(FCM)分析癌细胞DNA含量。结果 10 μg/mlpolα特异的ASODN在Lip 介导下可显著抑制人胃癌细胞的DNA复制和细胞增殖,正义ODN(SODN) 无此作用。结论 DNA- polα特异ASODN可用于胃癌的基因治疗, 阳离子脂质体可促进ASODN大量进入癌细胞, 增强其生物效应。
Objective To sutdy the growth inhibition of human gastric cancer cells by Lipofectin-mediated DNA polymerase α(DNA-pol α) antisense oligodeoxynucleotides (ASODN). Methods Oligonucleotides (ODN) including ASODN and sense ODN (SODN) targeted against the translation start site of pol α mRNA were synthesized and combined to Lipofectin (Lip) to prepare Lip+ODN complexes. After exposure to these complexes for 24 hours and continual incubation in complete medium for another 24 hours, the growth inhibition and DNA contents of MGC-803 and SGC-7901 human gastric cancer cells were studied by means of MTT assay and flow cytometry (FCM). Results 10 μg/ml of ASODN specific to pol α mediated by Lipofectin exhibited a significant effect on the DNA replication and proliferation of human gastric cancer cellos. Sense ODN (SODN) had no similar effect. Conclusions ASODN specific to pol α may be used in the gene therapy of cancer. Cationic liposome can increase the entry of ASODN into target cells and inprove the biological effect of ASODN.
出处
《广东医学》
CAS
CSCD
2000年第1期11-13,共3页
Guangdong Medical Journal