福多司坦对肺癌细胞黏蛋白MUC1和MUC5AC的抑制作用研究

Study of Inhibition of Fudosteine on Mucin MUC1 and MUC5AC in Lung Cancer Cells

关键词:  福多司坦  黏蛋白MUC1  黏蛋白MUC5AC  分子对接  细胞验证

Key words:  fudosteine  mucin MUC1  mucin MUC5AC  molecular docking  cell verification

摘要:

目的 探讨黏痰溶解剂福多司坦对肺癌细胞高表达的黏蛋白MUC1和MUC5AC的调控作用。方法 首先采用分子对接技术将福多司坦、原配体和阳性药与MUC1和MUC5AC蛋白进行对接，初步得到福多司坦、原配体及阳性药与靶蛋白的结合能与分子-蛋白相互作用活性对接口袋。然后采用体外PCR与ELISA从细胞层面探讨不同浓度福多司坦对TNF-a诱导的高表达MUC1和MUC5AC的调控作用。结果 分子对接结果显示福多司坦与MUC1和MUC5AC均能自发结合，并且福多司坦与MUC1的结合比MUC5AC更为紧密。细胞试验表明，不同浓度福多司坦(1~10 mmol·L^–1)可以降低肺癌A549细胞在TNF-a刺激下引起的MUC1和MUC5AC的高表达，也可降低NCI-H292细胞在TNF-a刺激下引起的MUC1的高表达。首次发现福多司坦能阻断肺癌细胞A549的MUC1蛋白表达，并且对MUC1蛋白的抑制呈现浓度依赖性。结论 福多司坦可抑制A549细胞黏蛋白MUC5AC和MUC1的表达，也可抑制NCI-H292细胞黏蛋白MUC1的表达，为进一步研究对肺癌细胞转移和侵袭的干预作用以及辅助肺黏液型肺癌患者的临床治疗提供理论依据。

Abstract:

OBJECTIVE To explore the regulatory effects of the mucoactive agent fudosteine on highly expressed mucins MUC1 and MUC5AC in lung cancer cells. METHODS Firstly, the molecular docking technology was used to dock fudosteine, the original ligand and the positive drug with MUC1 and MUC5AC proteins, and the binding energy value and the molecule-protein interaction activity of the docking pocket were preliminarily obtained. Then PCR and ELISA experiment were used to verify the regulatory effect of different concentrations of fudosteine on TNF-a-induced high expression of MUC1 and MUC5AC in vitro from the cellular level. RESULTS Molecular docking results showed that fudosteine spontaneously bound to MUC1 and MUC5AC, and that fodosteine binds more tightly to MUC1 than to MUC5AC. The results of cell verification showed that the high expression of MUC1 and MUC5AC in lung cancer A549 cells or the high expression of MUC1 in NCI-H292 cells stimulated by TNF-a could be inhibited by 1 mmol·L^-1 to 10 mmol·L^-1 of fudosteine, and the effect of lowering MUC1 protein expression was better than that of MUC5AC protein. Fudosteine was first found to block the expression of MUC1 protein in lung cancer A549 cells in a concentration-dependent way. CONCLUSION Fudosteine can inhibit the expression of MUC1 and MUC5AC in A549 and the expression of MUC1 in NCI-H292 lung cancer cell, and this study supplies a theoretical basis for intervening the metastasis and invasion of lung cancer cells and providing theoretical basis for the clinical treatment of patients with mucinous lung cancer.