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. Author manuscript; available in PMC: 2012 Dec 28.
Published in final edited form as: Oncogene. 2011 Nov 7;31(26):3111–3123. doi: 10.1038/onc.2011.498

Figure 3. Effect of CHI3L1 on migration and tube formation of endothelial cells.

Figure 3

(A) Representative photographs (upper panel) and mean numbers of migrated HUVECs (lower panel) at x200 magnification. SW480 cells (2 × 105) and HUVECs (1 × 104) were co-cultured for 4 hrs. Migration of HUVECs was significantly enhanced by co-culturing with SW480 cells (WT) than without SW480 cells (no cells). CHI3L1 stimulation (80 ng/ml) significantly enhanced the migration of HUVECs. Overexpression of CHI3L1 (CH) in SW480 cells significantly enhanced the migration of HUVECs as compared with empty vector (EV) transfection. The enhanced migration of HUVECs by CHI3L1 was significantly inhibited by anti-CHI3L1 neutralizing Ab (CH + α-CH Ab) as compared with control rabbit IgG (CH + IgG).

(B) Representative photographs (upper panel) and mean numbers of migrated HUVECs (lower panel) at x200 magnification. Two kinds of CHI3L1 miRNAs (miRNA 1 and miRNA 2) significantly suppressed the migration of HUVECs as compared with negative control miRNA (NC).

(C) Representative photographs (upper panel) and mean numbers of tube-forming structures (lower panel) at x100 magnification. HUVECs (5 × 104) were incubated in conditioned medium for 5 hrs. Tube formation of HUVECs was significantly enhanced by conditioned medium from SW480 cells (WT) as compared with basal medium. CHI3L1 stimulation (80 ng/ml) significantly enhanced the tube formation of HUVECs. Overexpression of CHI3L1 (CH) in SW480 cells significantly enhanced the tube formation as compared with empty vector (EV) transfection. The enhanced tube formation of HUVECs by CHI3L1 was significantly inhibited by treatment with anti-CHI3L1 neutralizing Ab (CH + α-CH Ab) as compared with control rabbit IgG (CH + IgG).

(D) Representative photographs (upper panel) and mean numbers of tube-forming structures (lower panel) at x100 magnification. Two kinds of CHI3L1 miRNAs (miRNA 1, miRNA 2) significantly suppressed the tube formation of HUVECs as compared with negative control miRNA (NC).

All results (A-D) are the mean (± SEM) of more than three separate experiments. NS, no significant difference, * P < 0.05, ** P < 0.01.