背景与目的肺癌细胞耐药已经成为肺癌化疗的主要困难之一，异长春花碱被认为可有效抑制肺癌细胞的增殖和转移。本研究旨在探讨异长春花碱对人肺癌A549/DDP细胞顺铂耐受性的逆转作用及机制。方法1μmol/L和5μmol/L异长春花碱作用A549/DDP细胞后，应用MTS法检测肿瘤细胞顺铂敏感性的变化，应用流式细胞术检测肿瘤细胞凋亡率变化，肿瘤细胞对Rh-123摄入量的变化，Western blot法检测MDR1、Bcl-2、survivin、caspase-3/8和PTEN蛋白表达以及Akt的磷酸化水平的变化，real-time PCR检测MDR1、Bcl-2、survivin和PTEN的mRNA表达，用报告基因系统检测NF-κB、Twist和Snail的转录活性。结果1μmol/L和5μmol/L异长春花碱作用A549/DDP细胞后，肿瘤细胞对顺铂的敏感性分别提高了1.91倍和2.54倍，肿瘤细胞对Rh-123的摄入量提高了1.93倍和2.95倍，细胞凋亡增加了2.25倍和3.82倍，MDR1、Bcl-2、survivin蛋白表达和Akt磷酸化水平下调，caspase-3/8和PTEN蛋白表达上调，MDR1的mRNA表达下调43.5%和25.8%，Bcl-2的mRNA表达下调57.3%和34.1%，survivin的mRNA表达下调37.6%和12.4%，PTEN表达上调183.4%和154.2%，NF-κB转录活性下降53.2%和34.5%，Twist转录活性下降61.4%和33.5%， Snail转录活性下降57.8%和18.7%。结论异长春花碱可提高肿瘤细胞A549/DDP对顺铂的敏感性，其机制可能与调节PTEN/AKT/NF-κB信号路径活性，进而下调耐药基因表达，上调促凋亡基因表达有关。%Background and objective Drug resistance is a major obstacle on lung cancer treatment and Vinorel-bine is an effective drug to inhibition of tumor proliferation and metastasis. In this study, we investigated the effect and mecha-nism of Vinorelbine on reversing the cisplatin resistance of human lung cancer A549/DDP cell line. Methods With 1μmol/L and 5μmol/L Vinorelbine treatment, MTS assay was employed to determine the effect of the cisplatin sensitivity of tumor cells, lfow cytometry to determine the apoptosis rate and change of Rh-123 content;Western blot to determine the expression of MDR1, Bcl-2, surviving, PTEN, caspase-3/8 and phosphorylation level of Akt (p-Akt);Real-time PCR was to determine the mRNA expression of MDR1, Bcl-2, survivin and PTEN. Finally the transcriptional activities of NF-κB, Twist and Snail were determined by reporter gene system. Results With 1μmol/L and 5μmol/L Vinorelbine treatment, the sensitivity of cancer cells to cisplatin was increased by 1.91-and 2.54-folds respectively, lfow cytometry showed that the content of Rh-123 was el-evated 1.93-and 2.95-folds and apoptosis rate was increased 2.25-and 3.82-folds, Western blot showed that the expression of multidrug resistance related proteins MDR, Bcl-2 and survivin were downregulated, caspase-3/8 and PTEN was upregulated, phosphorylation of Akt was downregulated as well, real-time assay showed that the mRNA expression of MDR1 was down-regulated 43.5%and 25.8%, Bcl-2 was downregulated 57.3%and 34.1%, survivin was downregulated 37.6%and 12.4%, PTEN was upregulated 183.4%and 154.2%, the transcriptional activities of NF-κB was downregulated 53.2%and 34.5%, Twist was downregulated 61.4%and 33.5%, and Snail was downregulated 57.8%and 18.7%. Conclusion Vinorelbine treatment led to increase of cisplatin sensitivity of A549/DDP cells and the mechanisms included the regulation of PTEN/AKT/NF-κB signal pathway to decreased drug resistance gene expression and increased pro-apoptosis gene expression.
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