LIU Bing, ZHANG Hong, LI Wenjian, et al. Effects of AdCMV-p53 gene transfer induced by irradiation on cycle of human colorectal adenocarcinoma cells[J]. Journal of Radiation Research and Radiation Processing, 2006,24(2):102-106.
LIU Bing, ZHANG Hong, LI Wenjian, et al. Effects of AdCMV-p53 gene transfer induced by irradiation on cycle of human colorectal adenocarcinoma cells[J]. Journal of Radiation Research and Radiation Processing, 2006,24(2):102-106.DOI:
以复制缺陷型重组腺病毒载体 (,AdCMV,-,GFP,)为对照,用复制缺陷型,p53,重组腺病毒载体 (,AdCMV,-,p53,)转染经0.5、1.0、2.0Gy γ射线照射的HT-29细胞,克隆形成法检测对细胞的抑制作用,流式细胞分析法检测细胞周期和细胞凋亡,探讨辐射诱导对,AdCMV,-,p53,转染,p53,突变型结直肠癌细胞(HT-29细胞系)细胞周期的影响。结果显示,0.5—1.0Gy辐射诱导明显增强40 MOI ,AdCMV,-,p53,转染对HT-29细胞的抑制。与,AdCMV,-,p53,转染对照相比,1d后,辐射诱导转染组G,0,/G,1,期细胞减少5%—15%,S期细胞增加2%—19%,2.0Gy辐射诱导80 MOI ,AdCMV,-,p53,转染组G,2,/M期细胞增加12%;3d后,0.5、1.0Gy辐射诱导40 MOI ,AdCMV,-,p53,转染组G,2,/M期细胞分别增加10%—13%。辐射诱导,AdCMV,-,p53,转染组细胞凋亡与辐射诱导剂量和,AdCMV,-,p53,转染剂量相关。以上结果表明,辐射诱导加速,AdCMV,-,p53,转染细胞由G,0,/G,1,期到S期的进程,促进S期阻滞和G,2,/M期阻滞发生。
Abstract
The work is to investigate effects of ,AdCMV-p53, gene transfer induced by ,60,Co γ-rays on cell cycles of human colorectal adenocarcinoma cells. HT-29 cells exposed to 0.5, 1.0 and 2.0Gy were infected with ,AdCMV-GFP, a replication deficient recombinant adenoviral vector containing a CMV promoter and green fluorescent protein, or ,AdCMV-p53, a replication deficient recombinant adenoviral vector containing a CMV promoter and carrying human wild-type ,p53, gene. Survival rate of the cells was determined by clonogenic assay. Cell cycle and cell apoptosis were determined by flow cytometry. The results showed that, 0.5—1.0Gy irradiation significantly enhanced the inhibition of ,AdCMV-p53, infection on HT-29. Compared with the control, 1day after the infection, the cells in G,0,/G,1, phase decreased by 5%—15%, the cells in S phase increased by 2%—19%. The 0.5 and 1.0Gy irradiation made the cells in the in G,2,/M phase increase by 12%, infected with 80 MOI ,AdCMV-p5,. Three days later, the proportion of cells in G,2,/M phase in groups of 0.5 and 1.0Gy irradiation + 40 MOI ,AdCMV-p53, infection increased by 10%—13%. There was a relation between cell apoptosis and irradiation dose, or ,AdCMV-p53, dose. Therefore, the irradiation-induction could quicken the progression from G,0,/G,1, phase to S phase, and promote S and G,2,/M phase arrest.