化疗药物对人肺腺癌细胞死亡受体DR4与DR5表达影响的研究

Study on effects of chemical drugs on expression of death receptors DR4 and DR5 in human lung adenocarcinoma cells

目的:研究化疗药物依托泊苷(Vp16)、顺铂(DDP)、多西他赛(TXT)对A549细胞死亡受体( death receptor, DR) DR4DR5基因表达的影响。方法:采用逆转录聚合酶链反应(PT PCR)方法对亚毒性剂量Vp 16 、DDP和TXT作用不同时间后的人肺腺癌细胞株A549 细胞的死亡受体DR4、DR5 的mRNA表达进行半定量检测。结果:1)亚毒性剂量Vp 16 可上调A549 细胞DR4、DR5 基因表达。7 5μg/mL Vp 16 处理A549 细胞8 h后, DR4/βactin、DR5/βactin 的比值分别为1. 13±0. 13 和1 .06±0. 25。7 5μg/mL Vp 16处理A549细胞12 h后,DR4/βactin、DR5/βac tin的比值分别为1 .18±0 .20和1.02±0. 02,两者与对照组之间比较,差异均有统计学意义,P<0 .05(P值分别为0 .04、0 02、0. 00和0 .02)2)亚毒性剂量DDP和TXT对A549细胞DR4DR5基因转录水平无明显影响。5μg/mLDDP、0 005μmol/L TXT (约为4 ng/mL) 作用A549细胞4、8 h后DR4、DR5基因转录水平与对照组之间比较,差异无统计学意义,P>0 .05结论:药物干预对TRAIL凋亡通路以及耐药机制的影响存在多样性。部分化疗药物可上调肿瘤细胞死亡受体DR4、DR5基因表达,同时不排除其他作用机制的存在。

OBJECTIVE:To study the effects of chemical drugs Vp-16,DDP and TXT on the gene expression of death receptors DR4 and DR5 in A549 cells. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was applied to semi-quantitatively assay mRNA expression of death receptors DR4 and DR5 in human lung adenocarcinoma cells A549 before and after the treatment of sub-toxic doses of Vp-16,DDP and TXT for different hours. RESULTS: 1)Sub-toxic dose of Vp-16 up-regulated the expression of DR4 and DR5 mRNA in A549 cells. The ratios of DR4/β-actin and DR5/β-actin in A549 cells treated by Vp-16(7.5 μg/mL) for 8 h were 1.13±0.13 and 1.06±0.25, and the ratios treated for 12 h 1.18±0.20 and 1.02±0.02. There were significantly statistical differences between the above two groups and normal control group, P < 0.05( P =0.04,0.02,0.00 and 0.02).2)Sub-toxic doses of DDP or TXT had no significant effects on the expression of DR4 and DR5 mRNA in A549 cells. The expression of DR4 and DR5 mRNA in A549 cells treated by DDP(5 μg/mL) or TXT(0.005 μmol/L) for 4 h and 8 h was not significantly higher than that of normal control groups, P >0.05. CONCLUSIONS: The effects of reversal by drugs on TRAIL apoptosis pathway and mechanisms of resistance to TRAIL are diverse. Some of chemical drugs can up-regulate the expression of death receptors DR4 and DR5 mRNA in tumor cells. At the same time, the possibility of existence of other mechanisms can not be excluded simply.