pEgr1-Trail重组质粒联合电离辐射对MCF-7细胞杀伤效应的实验研究

目的研究p Egr1-Trail重组质粒联合电离辐射对MCF-7细胞的杀伤效应,为增强肿瘤放疗效果提供实验证据.方法应用Annexin V-EGFP/PI双染法检测细胞凋亡;应用酶标仪检测细胞增殖;应用流式细胞仪检测细胞周期.结果不同处理组作用MCF-7细胞后24 h、不同剂量X射线照射后24 h后,各处理组MCF-7细胞的生长抑制率呈上升态势,生长抑制由弱到强的顺序为:2.0 Gy,p Egr1-Trail+0.5 Gy,5.0 Gy,p Egr1-Trail+1.0 Gy,p Egr1-Trail+2.0 Gy,Egr1-Trail+5.0 Gy.各处理组联合2.0 Gy X射线照射后6 h,细胞凋亡率开始上升,48 h达高峰,其中p Egr1-Trail+2.0 Gy组与其他各处理组比较差异具有统计学意义,且细胞凋亡最为显著(P0.05).在细胞周期方面,2.0 Gy X射线照后6 h各处理组S期细胞百分数呈现上升趋势,24 h达高峰;照后24 h,G_2+M期细胞百分数开始上升,且各处理组比较差异均具有统计学意义(P0.05);照后48 h,G_2+M期细胞百分数达到最高,依旧是p Egr1-Trail+2.0 Gy组上升最显著.结论 p Egr1-Trail重组质粒联合电离辐射作用于肿瘤细胞表现出协同杀伤效应,促凋亡作用强于单独X射照射组或p Egr1-Trail基因干预组.

Experimental Research on Killing Effects of pEgr1-Trail Recombinant Plasmid Combined with Ionizing Radiation on MCF-7 Cell

Objective To study the killing effects of pEgr1-Trail recombinant plasmid combined with ionizing radiation on MCF-7 cell,and to provide experimental evidence for improving curative effects of radiotherapy for tumor. Method Annexin V-EGFP/PI was used to detect cell apoptosis, cell proliferation was detected by ELIASA,and cell cycle was determined by flow cytometer. Results After being irradiated for 24 h by different doses of X-ray,the growth inhibition rates of MCF-7 cell in different treatment groups all increased with the order of 2. 0 Gy, pEgr1 Trail+0. 5 Gy,5. 0 Gy, pEgr1-Trail+1. 0 Gy, pEgr1-Trail+2. 0 Gy, pEgr1-Trail+5. 0 Gy. Compared with other groups,PEgr1-Trail group had significant difference (P<0. 05). In the combination of 2. 0 Gy X-ray irradiation for 6 h,apoptosis rates of each group started to rise,and reached peak at 48 h. Compared with other treatment group,cell apoptosis of pEgr1-Trail +2. 0 Gy group was the most obvious (P<0. 05). As for cell cycle,compared with the control group,cell percentage in S phase of each treatment group began to rise after being irradiated by 2. 0 Gy X-ray for 6 h,and got to peak at 12 h (P<0. 05). After 24 h,cell percentages in G2+M phase began to rise,and the differences among different treatment groups were all significant (P<0. 05). After 48 h,cell percentages in G2+M phase were the highest,and the percentage was aslo the most obvious in pEgr1-Trail+2. 0 Gy group. Conclusion PEgr1-Trail recombinant plasmid combined with ionizing radiation exhibit synergistic effect on killing tumour cells,their synergistic effect on promoting apoptosis is better than the single use of X-ray irradiation or gene interference.