title: Enoxaparin sensitizes human non-small-cell lung carcinomas to gefitinib by inhibiting DOCK1 expression, vimentin phosphorylation, and Akt activation.
abstract: Gefitinib is widely used for the treatment of lung cancer in patients with sensitizing epidermal growth factor receptor mutations, but patients tend to develop resistance after an average of 10 months. Low molecular weight heparins, such as enoxaparin, potently inhibit experimental metastasis. This study aimed to determine the potential of combined enoxaparin and gefitinib (enoxaparin + gefitinib) treatment to inhibit tumor resistance to gefitinib both in vitro and in vivo. A549 and H1975 cell migration was analyzed in wound closure and Transwell assays. Akt and extracellular signal-related kinase 1/2 signaling pathways were identified, and a proteomics analysis was conducted using SDS-PAGE/liquid chromatography-tandem mass spectrometry analysis. Molecular interaction networks were visualized using the Cytoscape bioinformatics platform. Protein expression of dedicator of cytokinesis 1 (DOCK1) and cytoskeleton intermediate filament vimentin were identified using an enzyme-linked immunosorbent assay, Western blot, and small interfering RNA transfection of A549 cells. In xenograft A549-luc-C8 tumors in nude mice, enoxaparin + gefitinib inhibited tumor growth and reduced lung colony formation compared with gefitinib alone. Furthermore, the combination had stronger inhibitory effects on cell migration than either agent used individually. Additional enoxaparin administration resulted in better effective inhibition of Akt activity compared with gefitinib alone. Proteomics and network analysis implicated DOCK1 as the key node molecule. Western blot verified the effective inhibition of the expression of DOCK1 and vimentin phosphorylation by enoxaparin + gefitinib compared with gefitinib alone. DOCK1 knockdown confirmed its role in cell migration, Akt expression, and vimentin phosphorylation. Our data indicate that enoxaparin sensitizes gefitinib antitumor and antimigration activity in lung cancer by suppressing DOCK1 expression, Akt activity, and vimentin phosphorylation.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Yan
authors.initials: Y
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Jianhui
authors.initials: J
authors.lastname: Duan
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Lan
authors.initials: L
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Shengjun
authors.initials: S
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Jingpu
authors.initials: J
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Yilixiati
authors.initials: Y
authors.lastname: Xiaokaiti
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Haopeng
authors.initials: H
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.).
authors.forename: Yefan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Health Science Center and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, People's Republic of China (Y.P., X.L., J.D., S.F., J.F., Y.X., H.Y., Y.W., X.L.); and Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing, People's Republic of China (L.Y.) xjli@bjmu.edu.cn.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000008:administration & dosage; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D017984:Enoxaparin / Q000008:administration & dosage*; D000077156:Gefitinib; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010766:Phosphorylation / Q000187:drug effects / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011799:Quinazolines / Q000008:administration & dosage*; D014746:Vimentin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays / Q000379:methods; D020744:rac GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis*
chemical_list: C099818:DOCK1 protein, human; D017984:Enoxaparin; D011799:Quinazolines; D014746:Vimentin; D051057:Proto-Oncogene Proteins c-akt; D020744:rac GTP-Binding Proteins; D000077156:Gefitinib