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目的观察血管内皮细胞生长因子反义寡核苷酸(VEGF-ASODN)转染胃癌细胞SGC-7901对血管内皮生长因子(VEGF)和生存素(survivin)表达的影响。方法人工合成硫代磷酸化VEGF-ASODN。实验分为5组:对照组,错义组,实验1组(转染浓度为100ng/mL),实验2组(转染浓度为400ng/mL)及实验3组(转染浓度为700ng/mL)。转染后实时荧光定量RT-PCR检测细胞RNA起始拷贝数;ELESA法检测细胞及培养上清液VEGF蛋白含量;Western-blot法检测细胞生存素蛋白表达量;流式细胞仪检测细胞凋亡;细胞生长曲线检测转染对细胞生长的影响。结果3个实验组VEGF mRNA水平和VEGF蛋白表达量及培养上清液VEGF蛋白含量均明显降低,survivin蛋白的表达水平随转染浓度的升高而降低,转染VEGF-ASODN能使胃癌细胞凋亡增加、细胞生长减缓。结论VEGF-ASODN转染胃癌细胞SGC-7901能显著抑制细胞VEGF的表达,促进细胞凋亡,抑制细胞增殖。
Objective To observe the effect of vascular endothelial growth factor antisense oligonucleotide (VEGF-ASODN) on the expression of vascular endothelial growth factor (VEGF) and survivin in gastric cancer cell line SGC-7901. Methods Synthesis of thiophosphorylated VEGF-ASODN. The experiment was divided into 5 groups: control group, missense group, experimental group 1 (100ng / mL), group 2 (400ng / mL) and group 3 ). Transfection of real-time fluorescent quantitative RT-PCR detection of the initial copy number of cellular RNA; ELESA assay of cell and culture supernatant VEGF protein content; Western-blot assay of cell survivin expression; flow cytometry apoptosis Cell growth curve was used to detect the effect of transfection on cell growth. Results The expression of VEGF mRNA, VEGF protein and VEGF protein in the supernatant of the three experimental groups were significantly decreased, while the expression of survivin protein decreased with the increase of the transfection concentration. The transfection of VEGF-ASODN could significantly inhibit the apoptosis of gastric cancer cells Increased death, slowed cell growth. Conclusion Transfection of SGC-7901 cells with VEGF-ASODN can significantly inhibit the expression of VEGF, promote cell apoptosis and inhibit cell proliferation.