腺病毒介导的CDglyTK基因影响胃癌细胞形态学及生长特性的效应

Influences of adenovirus-mediated CDglyTK gene on morphology and growth characteristics of gastric cancer cells

目的:观察已转染双自杀基因的胃癌细胞和未转染双自杀基因的胃癌细胞在形态学及生长特性方面的差异性。方法:实验于2005-03/2006-03在南方医科大学肿瘤研究所完成。质粒pAdEasy-KDR-CDglyTK为南方医科大学珠江医院普外科保存。用感染复数为100时,将启动子融合基因腺病毒(Ad-KDR-CDglyTK)体外感染SCG7901细胞株。通过倒置荧光相差显微镜、光镜、透射电镜等技术观察转染基因的胃癌细胞的形态学变化。通过流式细胞仪检测细胞周期的变化,同时用四甲基偶氮唑盐方法进行生长曲线的测定。结果:①腺病毒介导的KDR启动子驱动的双自杀基因对胃癌细胞体外培养的形态无明显影响。②已转基因的SCG7901细胞和未转基因的SCG7901细胞处于G1期,比率分别为(49.00±0.58)%,(51.02±0.91)%(t=1.862,P=1.12),处于S期比率分别为(16.20±0.36)%,(14.00±0.99)%(t=2.07,P=0.84),处于G2期比率分别为(34.60±0.96)%,(34.31±0.86)%(t=0.223,P=0.831)。③已转基因SCG7901细胞和未转基因SCG7901细胞具有类似的生长状态(F=0.042,P=0.838>0.05)。结论:已转染KDR启动子融合基因的胃癌细胞和未转染的胃癌细胞在形态及生长特性方面无显著性差异。

AIM： To observe the similarities and differences of the morphology and characteristics of growth between the transfected double suicide gene gastric cancer cells and non-transfected double suicide gene gastric cancer cells. METHODS： The experiment was conducted at the Tumor Institute of Southern Medical University from March 2005 to March 2006. Plasmid pAdEasy-KDR-CDglyTK was kept by Department of General Surgery, Zhujiang Hospital, Southern Medical University. The gastric cancer cells SCG7901 were infected by the Ad-KDR-CDglyTK in which multiplicity of infection （MOI） of Ads was 100 in vitro. The change of morphology of the transfected cancer cells were observed under inverted fluorescent contrast phase microscope, light microscope and transmission electron microscope. Moreover, the distribution of cell cycle was detected with flow cytometer and growth curve was measured with diphenyltetrazolium bromide （MTT） assay. RESULTS： （1）Double suicide gene had no significant effect on the formation of gastric cancer cells cultured in vitro drived by KDR promoter mediated by adenovirns. （2）The transfected SCG7901 cells and non-transfected SCG7901 cells were in G1 phase, and the ratio was （49.00±0.58）%, （51.02±0.91）%（t=l.862,P=-l.12）, and in S phase the ratio was （16.20±0.36）%, （14.00±0.99）%（t=2.07,P=0.84）, and in G2 phase the ratio was （34.60±0.96）%, （34.31 ±0.86）%（t=0.223,P=0.831）. （3）The transfected SCG7901 cells and non-transfected SCG7901 cells had the similar growth （F=0.042,P=-0.838 〉 0.05）. CONCLUSION： The significant difference have not been founded in the morphology and characteristics of growth both the transfected KDR promoter fusion gene gastric cancer cells and non-transfected gastric cancer cells.