Cellular and pharmacogenetics foundation of synergistic interaction of pemetrexed and gemcitabine in human non-small-cell lung cancer cells
Cellular and pharmacogenetics foundation of synergistic interaction of pemetrexed and gemcitabine in human non-small-cell lung cancer cells
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DOI：
10.1124/mol.104.009373
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发表时间：
2005-07-01
影响因子：
3.6
通讯作者：
Danesi, R
中科院分区：
文献类型：
--
作者：
Giovannetti, E;Mey, V;Danesi, R
吉西他滨和培美曲塞是治疗非小细胞肺癌 (NSCLC) 的有效药物，本研究调查了它们对 A549、Calu-1 和 Calu-6 细胞相互作用的细胞和遗传方面。用培美曲塞和吉西他滨处理细胞，并使用组合指数评估它们的相互作用。使用脱氧胞苷激酶（dCK）、5'-核苷酸酶和胞苷脱氨酶抑制剂检查了药物代谢在吉西他滨细胞毒性中的作用，而培美曲塞靶标、胸苷酸合酶（TS）、二氢叶酸还原酶（DHFR）和甘氨酰胺核糖核苷酸甲酰基转移酶（GARFT）在药物化疗敏感性中的作用进行了分析。细胞毒性救援研究。通过酶联免疫吸附测定研究了吉西他滨和培美曲塞对 Akt 磷酸化的影响，而定量聚合酶链反应 (PCR) 用于研究靶基因表达谱及其每种药物的调节。培美曲塞具有协同细胞毒性，可显着降低 A549 和 Calu-6 细胞中磷酸化 Akt 的量，增强细胞凋亡，增加 dCK 的表达，以及所有细胞系中人核苷平衡转运蛋白 1 (hENT1) 的表达。 PCR 证明 dCK 表达与吉西他滨敏感性之间存在相关性，而 TS、DHFR 和 GARFT 的表达可预测培美曲塞化疗敏感性。这些数据表明，1) 吉西他滨和培美曲塞通过抑制 Akt 磷酸化和诱导细胞凋亡，对 NSCLC 细胞产生协同作用； 2) 关键基因的基因表达谱可以预测药物化疗敏感性； 3) 培美曲塞增强 dCK 和 hENT1 表达，从而表明基因表达调节对于合理开发化疗组合的作用。
Gemcitabine and pemetrexed are effective agents in the treatment of non - small-cell lung cancer (NSCLC), and the present study investigates cellular and genetic aspects of their interaction against A549, Calu-1, and Calu-6 cells. Cells were treated with pemetrexed and gemcitabine, and their interaction was assessed using the combination index. The role of drug metabolism in gemcitabine cytotoxicity was examined with inhibitors of deoxycytidine kinase (dCK), 5'-nucleotidase, and cytidine deaminase, whereas the role of pemetrexed targets, thymidylate synthase (TS), dihydrofolate reductase ( DHFR), and glycinamide ribonucleotide formyltransferase ( GARFT) in drug chemosensitivity was analyzed in cytotoxicity rescue studies. The effect of gemcitabine and pemetrexed on Akt phosphorylation was investigated with enzyme-linked immunosorbent assay, whereas quantitative polymerase chain reaction (PCR) was used to study target gene-expression profiles and its modulation by each drug. Synergistic cytotoxicity was demonstrated, and pemetrexed significantly decreased the amount of phosphorylated Akt, enhanced apoptosis, and increased the expression of dCK in A549 and Calu-6 cells, as well as the expression of the human nucleoside equilibrative transporter 1 (hENT1) in all cell lines. PCR demonstrated a correlation between dCK expression and gemcitabine sensitivity, whereas expression of TS, DHFR, and GARFT was predictive of pemetrexed chemosensitivity. These data demonstrated that 1) gemcitabine and pemetrexed synergistically interact against NSCLC cells through the suppression of Akt phosphorylation and induction of apoptosis; 2) the gene expression profile of critical genes may predict for drug chemosensitivity; and 3) pemetrexed enhances dCK and hENT1 expression, thus suggesting the role of gene-expression modulation for rational development of chemotherapy combinations.