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探讨了辐射诱导对p53重组腺病毒载体转染p53突变型结直肠癌细胞(HT-29细胞系)的影响。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、2.0Gyγ射线照射的HT-29细胞,用流式细胞分析法检测细胞凋亡和p53蛋白表达,并用克隆形成法测定细胞增殖能力。结果表明,0.5Gy辐射诱导明显提高了AdCMV-p53转染对HT-29细胞的抑制,促进细胞凋亡。0.5Gy辐射诱导对AdCMV-p53转染HT-29细胞的抑制杀伤的增强作用最强,0.5Gy辐射诱导及40MOI和80MOIAdCMV-p53转染对肿瘤的抑制率比同等剂量下单纯转染组分别提高约50%和20%,比0.5Gy单纯辐照组提高40%。因此,0.5Gy辐射诱导可有效增加低于80MOIAdCMV-p53转染对HT-29细胞的抑制作用。辐射诱导AdCMV-p53转染HT-29细胞以最大辐射剂量不超过1.0Gy及AdCMV-p53转染量低于80MOI为宜。
The effects of radiation-induced p53 recombinant adenovirus vector transfection on p53 mutant colorectal cancer cells (HT-29 cell line) were investigated. The replication-defective recombinant adenovirus vector (AdCMV-GFP) was used as a control, and HT-29 cells irradiated with 0.5, 1.0, and 2.0 Gy γ-rays were transfected with the replication-defective p53 recombinant adenoviral vector (AdCMV-p53). The cell apoptosis and p53 protein expression were detected by cell assay, and cell proliferation ability was determined by colony formation assay. The results showed that 0.5Gy irradiation significantly enhanced the inhibition of AdCMV-p53 transfection on HT-29 cells and promoted apoptosis. The 0.5Gy radiation induced the strongest inhibitory effect on the inhibition of AdCMV-p53 transfected HT-29 cells. The inhibition rate of 0.5Gy and 40MOI and 80MOIAdCMV-p53 transfected tumors was higher than that of the single transfection group at the same dose. About 50% and 20%, 40% higher than 0.5Gy pure irradiation group. Therefore, 0.5Gy radiation induction can effectively increase the inhibitory effect of 80MOIAdCMV-p53 transfection on HT-29 cells. Radiation-induced transfection of HT-29 cells with AdCMV-p53 was not more than 1.0 Gy and AdCMV-p53 transfection was less than 80 MOI.