TRAIL基因修饰联合阿霉素对大肠癌细胞株RKO作用的研究

目的:探讨肿瘤坏死因子(tumor necrsis factor,TNF)相关的凋亡诱导配体(TNF-related apoptosis-inducing ligand,TRAIL)基因联合阿霉索后,应用于人大肠癌细胞株RKO基因治疗的实验研究.方法:将重组腺病毒载体(Ad)介导的TRAIL基因作用于大肠癌细胞株RKO,并联合低剂量的阿霉素协同作用.通过MTF比色法与流式细胞仪研究分析其对RKO细胞的作用效果,并以RT-PCR检测联合应用阿霉素前后TRAIL基因的表达水平.结果:病毒载体对RKO细胞的生长有轻微的抑制作用,作用4 d抑制率为11.9%,但不增加RKO细胞的凋亡率.TRAIL对RKO细胞的生长抑制率及凋亡诱导率分别为50.1%和19.8%.联合阿霉素后,TRAIL对RKO细胞株的生长抑制率及凋亡率均有显著的增强作用,分别达60.3%及49.0%.RT-PCR结果提示联合应用阿霉素后,TRAIL基因的表达并未增强.结论:TRAIL能有效抑制RKO的生长,联合阿霉素后,其对RKO的生长抑制作用及凋亡诱导作用均明显增强.阿霉素不是通过增加TRAIL基因的表达来实现上述作用的.

Activity of TRAIL Gene Modification Combined with DOX on Human Colon Cancer Cell Line RKO

Objective: To evaluate the gene therapeutic efficiency of TRAIL combined with DOX on human colon cancer cell line RKO. Methods: Human colon cancer cell line RKO was transfected with adenovirus-mediated TRAIL gene Ad/GT-TRAIL and TRAIL/DOX combination. Cell growth and apoptosis were measured by MTT method and flow cytome-try. TRAIL gene expression before and after combination with DOX was measured by RT-PCR. Results: The cell line RKO was slightly suppressed by mediate adenovirus and significantly suppressed by TRAIL gene. The suppression percentages were 11.9% and 50. 1% respectively. Combined with DOX, the suppression and the apoptosis of cell line RKO could be significantly enhanced by TRAIL gene. The suppression and the apoptic percentage reached 60. 3% and 49.0% respectively. RT-PCR showed the expression of TRAIL gene was not enhanced in the RKO cells exposed to the DOX/ TRAIL combination. Conclusion: TRAIL gene was able to induce apoptosis and suppress the growth of human colon cancer cell line RKO. Combined with DOX, the suppression and apoptosis could be significantly enhanced. But this result came out not by increasing the expression of TRAIL gene.