Lipofectin介导DNA聚合酶α反义寡核苷酸抑制人胃癌细胞生长

The growth inhibition of human gastric cancer cells by Lipofection-mediated of DNA polymerase alpha antisense oligodeoxynucleotides

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.