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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 study the inhibitory effect of cationic liposome Lipofectin (Lip) -mediated DNA polymerase α (ASODN) on the growth of human gastric cancer cells. Methods An oligonucleotide (ODN) complementary to 18 nucleotides of translation initiation region of human pol α catalytic subunit was synthesized and mixed with Lip to prepare Lip + ODN complex. The ODN complex was applied to MGC-803 and SGC-7901 human gastric cancer cells 24 h, the cells were replaced and continue to culture for 24 h. Cell viability was measured by MTT colorimetric assay. The DNA content of cancer cells was analyzed by flow cytometry (FCM). Results 10 μg / ml polα-specific ASODN could significantly inhibit the DNA replication and cell proliferation of human gastric cancer cells under the action of Lip, and no effect of ODN (SODN). Conclusion DNA-pol α-specific ASODN can be used in gene therapy of gastric cancer. Cationic liposomes can promote ASODN into large numbers in cancer cells and enhance its biological effects.