重组质粒pEgr-1-TRAIL辐射诱导乳腺癌细胞效应的实验研究

目的将具有辐射诱导表达特性的重组质粒p Egr-1-TRAIL转入乳腺癌MCF-7细胞中,诱导肿瘤杀伤基因TRAIL的表达,实现辐射和基因对MCF-7细胞的双重杀伤效应.方法用ELISA法检测不同剂量X射线诱导TRAIL表达的量效关系及2 Gy X射线照射后不同时间TRAIL的表达;用流式细胞仪检测不同处理组MCF-7细胞早期凋亡情况.结果不同剂量X射线照射转染p Egr-1-TRAIL重组质粒的乳腺癌细胞MCF-7,TRAIL表达量明显高于假照组(P0.001~0.05),其中5 Gy X射线照后表达量最高,TRAIL表达量为假照组的5.1倍.2 Gy X射线照射后4 h,TRAIL表达量明显升高(P0.05),并随时间延长逐渐增加,照射后32 h达峰值,表达量为照射前的4.6倍.MCF-7-p Egr-1-TRAIL/0 Gy组早期凋亡细胞百分数较MCF-7/0 Gy和MCF-7-p3.1Egr/0 Gy组明显增加(P0.01),随着照射剂量的增加,MCF-7-p Egr-1-TRAIL/5 Gy组细胞早期凋亡率与其他处理组比较均存在统计学意义(P0.001~0.05),MCF-7/0 Gy组细胞生长最快,而MCF-7-p Egr-1-TRAIL/8 Gy组细胞生长最慢.结论乳腺癌细胞转染p Egr-1-TRAIL重组表达质粒联合X射线照射能够增加MCF-7细胞凋亡,抑制其生长.

Experimental Study on the Radiation Effects of Recombinant Plasmid pEgr-1-TRAIL for Breast Cancer Cells

Objective To induce the expression of TRAIL genes by transfering recombinant plasmid pEgr-1-TRAIL into breast cancer MCF-7 cells,and to implement killing effects of radiation and gene on MCF-7 cells. Method The concentration-response relationship of TRAIL expression induced by different doses of X-ray and the expression of TRAIL at different times under 2 Gy X-ray irradiation were detected with ELISA method. Early apoptosis of MCF-7 cells transferred by pEgr-1-TRAIL after different doses of X-ray irradiation was tested by flow cytometry. Results Breast cancer MCF-7 cells were transferred by recombinant plasmid pEgr-1-TRAIL under different doses of X-ray irradiation. The amounts of TRAIL expression were significantly higher than those of other groups (P<0. 001 ~0. 05). The highest expression amount was under 5 Gy X-ray,which was 5. 1 times of the control group. Under 2 Gy X-ray irradiation for 4 h,the expression amount of TRAIL increased significantly (P<0. 05),and reached the peak till 32 h,which was 4. 6 times of the control group. The percentage of early cell apoptosis of MCF-7-pEgr-1-TRAIL/0 Gy group increased obviously than that of MCF-7/0 Gy group and MCF-7-p3. 1 Egr/0 Gy group (P<0. 01). With the increase of irradiation doses,cell early apoptosis rate of MCF-7-pEgr-1-TRAIL/5 Gy group had significant difference compared with other treatment groups (P<0. 001 ~0. 05). Cells in MCF-7/0 Gy group grew the fastest,and the cell proliferation in MCF-7-pEgr-1-TRAIL/8 Gy group was the slowest. Conclusion Transfection of recombinant plasmid pEgr-1-TRAIL jointing X-ray irradiation can increase cell apoptosis and inhibit the growth of breast cancer MCF-7 cell.