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目的:观察尼古丁对胆管癌细胞QBC939化疗敏感性影响,并初步探讨其作用靶点。方法:应用MTT法检测α7烟碱型胆能受体激动剂尼古丁及其阻断剂α银环蛇毒素(α-BTX)干预后的人胆管癌细胞QBC939经5-FU处理后存活增殖能力变化,用单克隆平板试验观察尼古丁及α银环蛇毒素对5-FU处理后细胞克隆形成率变化。结果:经5-fu处理后,尼古丁刺激组(终浓分别为10-3g/L、10-4g/L、10-5g/L)细胞存活率分别为128%、124%、118%,细胞存活率较阴性对照组明显升高,并呈一定的浓度赖性,而α银环蛇毒素刺激组(2 ug/mL)、尼古丁α银环蛇毒素联合组细胞存活率分别为92%、94%、93%、92%。尼古丁刺激(6.2±0.40)的克隆形成能力明显高于α银环蛇毒素刺激组(3.2±0.20)、联合组(3.2±0.20)及对照组(3.4±0.33)。结论:尼古丁明显降低胆管癌细胞对化疗药物的敏感性,其可能是通过α7烟碱型胆碱能受体发挥化疗抵抗效应。
OBJECTIVE: To observe the effect of nicotine on the chemosensitivity of cholangiocarcinoma QBC939 cells and to explore its target. Methods: The viability and proliferation of human cholangiocarcinoma QBC939 cells treated with 5-FU were determined by MTT assay after nicotine α7 nicotinic agonist nicotine and its antagonist α-BTX The changes of cell clone formation rate after 5-FU treatment with nicotine and α-bungarotoxin were observed by monoclonal plate test. Results: After 5-fu treatment, the survival rates of cells in nicotine stimulation group (final concentrations 10-3g / L, 10-4g / L, 10-5g / L) were 128%, 124%, 118% Survival rate was significantly higher than the negative control group, and a certain concentration of dependence, and α bungarotoxin stimulation group (2 ug / mL), nicotine α bungarotoxin combined group cell survival rates were 92%, 94 %, 93%, 92%. The colony-forming ability of nicotine stimulation (6.2 ± 0.40) was significantly higher than that of α-bungarotoxin stimulation group (3.2 ± 0.20), the combination group (3.2 ± 0.20) and the control group (3.4 ± 0.33). Conclusion: Nicotine significantly reduces the sensitivity of cholangiocarcinoma cells to chemotherapeutic drugs, which may play a chemoresistance effect through α7 nicotinic cholinergic receptors.