| Background and ObjectiveLung cancer is the most common cancer and the most harmful to human’s health, with morbidity and mortality at the top of malignant tumors, and incidence rates are still rising year by year. Non-small cell lung cancer accounts for about80%of the total number of lung cancers. About70-80%patients with non-small cell lung cancer were diagnosed in advanced stage when they went to clinic at the first time for lung cancer related complications. About30-70%patients with early stage lung cancer, will relapse in focal or occur distance metastasis after surgical treatment. Chemotherapy is still one of the most important ways for treatment of advanced lung cancer. Although optimal chemotherapies are being founded for advanced non-small cell lung cancer, the treatment results are still not satisfactory. Therefore, new treatment technologies are still needed for non-small cell lung cancers. Some clinical researches have shown that molecular targeting drug combined with chemotherapy could improve the efficacy. The addition of bevacizumab to paclitaxel plus carboplatin in the treatment of selected patients with non-small-cell lung cancer has a significant survival benefit. Sunitinib is a multi-targeting antitumor agent, whose main mechanism is to inhibit vascular endothelial growth factor receptor. Sunitinib combined with chemotherapy can improve the curative efficacy in lung cancer patients. In our project we observe apoptosis in A549cell line, and also gene expression levels of c-Met, mek and erk by administration of sunitinib and/or docetaxel in different combination styles or time courses. Our study will investigate the more effective administration fomula of the above agents and provided a theoretical foundation of lung cancer therapy.Materials and Methods1. Human A549lung cell line was chosen for in vitro experiment.2. Sunitinib and docetaxel were two agents in this project.3. MTT assay was used to observe the suppressive effect of cell growth on A549cell, and half inhibitory concentration (half maximal inhibitory concentration, IC5o) was calculated.4. The flow cytometry (FCM) was used to analyze the alteration of cell cycle and cell apoptosis in A549cells.5. The alternations of mRNA expression levels of c-Met, mek and erk were detected by RT-PCR.6. Data were analyzed using SPSS18.0, P<0.05were taken as significant difference, and P<0.01showed the difference was very significant.Results1. The IC50values of sunitinib for A549cell line were (7.72±0.63),(5.93±0.76), and (3.81±0.48)μmol/L at24,48and72hours, and docetaxel were (29.03±0.52),(21.34±0.84) and (15.3±0.79)μg/ml.2. Efficacy of combination therapy with sunitinib malate and docetaxel was better than any of single agent. Combined treatment showed a significant correlation with the administration order. Obvious synergistic effect could be detected when using docetaxel before sunitinib, while not obvious effect be observed when using sunitinib before docetaxel or using sunitinib and docetaxel at the same time.3. The FCM results showed that A549cell number in the S stage was increased after sunitinib treatment, and G2/M cells increased by docetaxel treatment. When using docetaxel before sunitinib, cell number in S stage was obviously increased, and that in G0/G1stage reduced significantly.4. RT-PCR result shows that, when using docetaxel before sunitinib, the mRNA expression of c-Met, mek was downregulated and erk upregulated significantly.ConclusionsSunitinib and docetaxel could inhibit the growth of A549cell. The way using docetaxel before sunitinib shows obvious synergistic effect. |
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