重组TRAIL诱导肺腺癌细胞系细胞凋亡的实验研究

Apoptosis Study of Lung Cancer Cell Line A549 Treated by rsTRAIL

目的研究重组可溶性TRAIL作用于人肺癌系A549细胞培养物后,细胞凋亡的形态变化和凋亡率测定。方法MTT比色法测定重组TRAIL对肺腺癌A549细胞的生长抑制率。TRAIL处理前后细胞核、质的形态学变化。经不同浓度重组TRAIL处理后A549细胞的双色荧光变化。定量重组TRAIL与亚毒性浓度的CDDP联合使用,对肺腺癌A549细胞生长抑制率和凋亡率的影响。结果重组TRAIL对A549细胞生长有明显的抑制作用,TRAIL100μg.L-1作用24h后,抑制率达到41.6%。细胞形态学观察显示,A549肺腺癌细胞经50μg.L-1TRAIL处理24h后,可观察到典型的细胞凋亡特点。3.3mg.L-1的CDDP与50μg.L-1重组TRAIL合用,A549细胞凋亡率高达75.2%。结论重组TRAIL具有明显的诱导多种恶性肿瘤细胞凋亡的作用;化疗药物CDDP能加强重组TRAIL的抗肿瘤活性。

OBJECTIVE To study the apoptosis-induced effects of the recombinant TRAIL on lung cancer cell line A549, and investigate the combined anti-tumor effect of the recombinant TRAIL and CDDP. METHODS The morphology of cultured cell treated with or without the recombinant TRAIL were observed and compared microscopically. The anti-proliferative effects of the recombinant TRAIL on cancer cell line A549 was measured by MTF spectrophotometry method. The apoptosis rate of cell line A549 induced by different content of the recombinant TRAIl, were determined by AO/EB dual fluorescence stain followed by image analysis. To explore the methods to increase the sensitivity to TRAIL, cancer cell line A549 was treated with a lower dosage recombinant TRAIL and sub-toxici- ty level CDDP, and the combination of the anti-tumorigenesis effect determined by MTT spectrophotometry assay and flow cytometry were compared with the control groups treated with single TRAIL or CDDP. RESULTS The recombinant TRAIl. had significant antiproliterative effects on cancer cell line A549 treated with the TRAIL 100 μg·L^-1 for 24 h, the anti-proliferative rate of the cell line was 41.6%. Cells killed by the recombinant TRAIL showed typical apoptosis morphology changes. The apoptosis rate of the cells was also related to the level of the TRAIL Treated with 50 μg · L^-1 for 24 h, the apoptosis of A549 was obtained. The sub-toxicity level CDDP can significantly increased the sensitivity of A549. Treated with CDDP 3 mg · L^-1 and TRAIL 50 μg · L^-1, the apoptosis rate of A549 reached to 75.2%. CONCLUSION The Recombinant TRAIL can kill A549 cells by inducing cell apoptosis; CDDP can synergistically enhance the effect of the recombinant TRAIL on inducing apoptosis in A549 cells. The recombinant TRAIL is a promising anti-tumorgenesis drug, and the combination with CDDP may be an effective therapy strategy.