title: Orally Administered Mucolytic Drug l-Carbocisteine Inhibits Angiogenesis and Tumor Growth in Mice.
abstract: Angiogenesis, the formation of new blood vessels from pre-existing vessels, is essential for the growth and metastasis of tumors. In this study, we found that l-carbocisteine, a widely used expectorant, potently inhibits angiogenesis in vitro and in vivo. An in vivo Matrigel plug assay revealed that l-carbocisteine (2.5 mg/kg i.p. twice daily) significantly inhibited vascular endothelial growth factor (VEGF)-induced angiogenesis. l-Carbocisteine also suppressed VEGF-stimulated proliferation, migration, and formation of capillary-like structures of human umbilical vein endothelial cells (HUVECs). We examined the signaling pathways affected in VEGF-stimulated HUVECs, and found that l-carbocisteine significantly inhibited VEGF-induced phosphorylation of phospholipase C (PLC) γ, protein kinase C (PKC) μ, and extracellular signal-related kinases (ERK) 1/2, which have been shown to be essential for angiogenesis. However, these inhibitory effects of l-carbocisteine were not observed in the HeLa human cervical cancer cell line. An in vivo study of Colon-26 tumor-bearing mice found that tumor volumes were significantly smaller in mice treated with l-carbocisteine (150 mg/kg administered orally twice daily) in comparison with vehicle-treated mice. However, l-carbocisteine had no direct effect on Colon-26 cell proliferation or ERK activation. Collectively, our results suggest that l-carbocisteine inhibits tumor angiogenesis by suppressing PLCγ/PKC/ERK signaling.
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Shinya
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Yokota
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Shiori
authors.initials: S
authors.lastname: Nakayama
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Sayuri
authors.initials: S
authors.lastname: Oki
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Junpei
authors.initials: J
authors.lastname: Mutoh
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.).
authors.forename: Satoru
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Department of Clinical Biochemistry (T.S.,T.Y., S.N., S.O., K.S.) and Second Department of Pharmacology (J.M.), School of Pharmaceutical Science, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan; and Department of Immunobiology, School of Pharmacy and Pharmaceutical Science, Mukogawa Women's University, Nishinomiya, Hyogo, Japan (S.T.) satokei@phoenix.ac.jp.
authors.forename: Keizo
authors.initials: K
authors.lastname: Sato
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002233:Carbocysteine / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D005100:Expectorants / Q000494:pharmacology; D006367:HeLa Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000378:metabolism; D051966:Phospholipase C gamma / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D005100:Expectorants; D042461:Vascular Endothelial Growth Factor A; D002233:Carbocysteine; C089212:protein kinase D; D011493:Protein Kinase C; D051966:Phospholipase C gamma