5-Fu对人结肠癌细胞凋亡过程中DNA损伤修复基因表达的影响

Influence of 5-Fu on Expression of DNA Damage-Repair Gene During Apoptosis of Human Colon Cancer Cells

目的探讨5-Fu对人结肠癌细胞(HCT/8)凋亡过程中DNA损伤修复相关基因表达的影响。方法通过形态学、基因组电泳、吖啶橙染色及流式细胞仪,观察大肠癌细胞凋亡情况,同时应用基因芯片分析5-Fu作用后DNA损伤修复相关基因表达的变化。结果经流式细胞仪检测发现,5-Fu作用于HCT/8细胞48h与对照组相比,凋亡细胞数明显增加,基因组电泳出现典型的梯形条带,基因芯片分析显示,12个与DNA损伤修复相关的基因中,上调基因3个,下调基因9个。结论5-Fu作用于结肠癌细胞,导致多种DNA损伤修复基因发生改变,综合作用使细胞不能及时对损伤DNA进行修复,最终导致细胞发生凋亡。

Objective To explore the influence of 5-Fu on the expression of gene related to the repairing of DNA damage during the apoptosis of human colon cancer HCT/8 cells. Methods Induce the apoptosis of HCT/8 cells with 5-Fu and observe the morphological change of the cells. Determine the apoptosis of HCT/8 cells by genome electrophoresis, acridine orange staining and flow cytometer, and analyze the expression of gene related to the repairing of DNA damage by gene chip technique. Results The number of apoptotic cells 48 h after induction with 5-Fu increased significantly as compared with that of control cells, and typical ＂bladder bands＂ were shown on genome electrophoretic profile. The analysis by gene chip technique showed that the expression of 3 of the 12 genes related to the repairing of DNA damage was up-regulated, and that of the rest 9 genes were down-regulated. Conclusion 5-Fu induced the changes of many genes related to the repairing of DNA damage of human colon cancer cells, which could not repair the DNA damage in time and caused the cell apoptosis finally.