转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响  

Effect of ASODN transfection targeting VEGF-C gene on proliferation and apoptosis of Hela cells

作　　者：祁丽[1] 邢丽娜[1] 

机构地区：[1]哈尔滨医科大学附属第一医院肿瘤放疗科,150086

出　　处：《肿瘤研究与临床》2009年第7期437-439,443,共4页Cancer Research and Clinic

基　　金：哈尔滨市攻关课题（2005AA9CS116-10）

摘　　要：目的体外研究脂质体介导转染血管内皮生长因子C（VEGF—C）反义寡核苷酸（VEGF—C，ASODN）对子宫颈癌Hela细胞增生和凋亡的影响。方法实验组将VEGF—CASODN（优化筛选）用脂质体导人Hela细胞，设正义（SODN）和空白对照组进行比较。用荧光显微镜观察转染率，MTT比色法检测细胞增生，流式细胞术检测细胞凋亡及细胞周期变化，RT—PCR、Western blotting法检测VEGF—C mRNA和蛋白的表达。结果经脂质体介导可成功将VEGF—C反义核酸转染至子宫颈癌Hela细胞；细胞的增生受到明显抑制，G2／M期细胞比例增多，转染作用48h时凋亡率最高，达（19．39±1．81）％；VEGF—C在mRNA和蛋白水平的表达均明显减少，与对照组比较差异有统计学意义（P〈0．05）。结论体外转染VEGF—CASODN可在mRNA和蛋白水平下调Hela细胞VEGF—C的表达，阻滞并同步化细胞周期，抑制细胞增生，诱导细胞凋亡。Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C（VEGF-C） antisense oligonucleotides （ASODN）. Methods VEGF- C ASODN was transfected into Hela cells by liposome-mediated; the ceils transfected with the oligodeoxynuclecotide （SODN） and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was （19.39±1.81）%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein （P 〈0.05）. Conclusion Transfected with VEGF-C ASODN could downregulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.

关 键 词：子宫颈肿瘤 Hela细胞 血管内皮生长因子C 寡核苷酸类 反义 转染 细胞凋亡 

分 类 号：R737.33[医药卫生—肿瘤]