title: The M2 macrophages induce autophagic vascular disorder and promote mouse sensitivity to urethane-related lung carcinogenesis.
abstract: Tumor vessels are known to be abnormal, with typically aberrant, leaky and disordered vessels. Here, we investigated whether polarized macrophage phenotypes are involved in tumor abnormal angiogenesis and what is its mechanism. We found that there was no difference in chemotaxis of polarized M1 and M2 macrophages to lewis lung carcinoma (LLC) cells and that either M1 or M2 macrophage-conditioned media had no effect on LLC cell proliferation. Unexpectedly, the M2 but not M1 macrophage-conditioned media promoted the proliferation of human umbilical vein endothelial cells (HUVECs) and simultaneously increased endothelial cell permeability in vitro and angiogenic index in the chick embryo chorioallantoic membrane (CAM). The treatment with M2 but not M1 macrophage-conditioned media increased autophagosomes as well as microtubule-associated protein light chain 3B (LC3-B) expression (a robust marker of autophagosomes) but decreased p62 protein expression (a selective autophagy substrate) in HUVECs, the treatment with chloroquine that blocked autophagy abrogated the abnormal angiogenic efficacy of M2 macrophage-conditioned media. These results were confirmed in urethane-induced lung carcinogenic progression. Urethane-induced lung carcinogenesis led to more M2 macrophage phenotype and increased abnormal angiogenesis concomitant with the upregulation of LC3-B and the downregulation of p62. Clodronate liposome-induced macrophage depletion, chloroquine-induced autophagic prevention or salvianolic acid B-induced vascular protection decreased abnormal angiogenesis and lung carcinogenesis. In addition, we found that the tendency of age-related M2 macrophage polarization also promoted vascular permeability and carcinogenesis in urethane carcinogenic progression. These findings indicate that the M2 macrophages induce autophagic vascular disorder to promote lung cancer progression, and the autophagy improvement represents an efficacious strategy for abnormal angiogenesis and cancer prevention.
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: G-G
authors.initials: GG
authors.lastname: Li
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: Z-Z
authors.initials: ZZ
authors.lastname: Guo
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: X-F
authors.initials: XF
authors.lastname: Ma
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: N
authors.initials: N
authors.lastname: Cao
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: S-N
authors.initials: SN
authors.lastname: Geng
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: Y-Q
authors.initials: YQ
authors.lastname: Zheng
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: M-J
authors.initials: MJ
authors.lastname: Meng
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China.
authors.forename: H-H
authors.initials: HH
authors.lastname: Lin
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China. Electronic address: hang@henu.edu.cn.
authors.forename: G
authors.initials: G
authors.lastname: Han
authors.affiliation: Institute of Pharmacy, Pharmacy College of Henan University, Jinming District, Kaifeng, Henan Province 475004, China. Electronic address: 724200@henu.edu.cn.
authors.forename: G-J
authors.initials: GJ
authors.lastname: Du
mesh_terms: D000230:Adenocarcinoma / Q000139:chemically induced / Q000276:immunology*; D000077192:Adenocarcinoma of Lung; D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D001343:Autophagy / Q000276:immunology; D001572:Benzofurans / Q000494:pharmacology; D002199:Capillary Permeability / Q000502:physiology; D063646:Carcinogenesis / Q000139:chemically induced / Q000276:immunology*; D018827:Carcinoma, Lewis Lung; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002642:Chick Embryo; D002738:Chloroquine / Q000494:pharmacology; D004002:Clodronic Acid / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D042783:Endothelial Cells; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008175:Lung Neoplasms / Q000139:chemically induced / Q000276:immunology*; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis; D009389:Neovascularization, Pathologic / Q000276:immunology*; D010750:Phosphoproteins / Q000096:biosynthesis; D015534:Trans-Activators / Q000096:biosynthesis; D014520:Urethane / Q000494:pharmacology; D053630:Depsides
chemical_list: D001572:Benzofurans; D002738:Chloroquine; D004002:Clodronic Acid; D017077:Culture Media, Conditioned; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D015534:Trans-Activators; D014520:Urethane; C480860:Map1lc3b protein, mouse; C486419:Trp63 protein, mouse; C076944:salvianolic acid B; D053630:Depsides
keywords: Autophagy; Lung cancer; Macrophage polarization; Vascular disorder