摘要
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 effects of VEGF antisensc oligonucleotide transfected to SGC-7091 cell line on expression of VEGF and survivin of the cell. Methods The VEGF-ASODN was synthesised artificially. There were five groups in this experiment： control group, missense group and three various concentration groups. After transfection, RNA copies were detected by real-time PT-PCR, VEGF protein in cell and culture supemate was detected by ELESA, survivin protein was measured by Western-blot, and apoptosis was detected by FCM. Growth of cells was evaluated by growth curve. Results VEGF-ASODN tranfection reduced the VEGFmRNA and VEGF protein expression in gastric cancer cells and supernate remarkably. It also reduced survivin protein expression and increased apoptosis in gastric cancer cells. Conclusions Transfection with VEGF-ASODN to gastric cancer cells SGC-7901 can reduce the expression of VEGF, and can increase apoptosis and suppress the proliferation of gastric cancer cells.
出处
《中国普通外科杂志》
CAS
CSCD
2008年第10期983-987,共5页
China Journal of General Surgery
