Axl glycosylation mediates tumor cell proliferation, invasion and lymphatic metastasis in murine hepatocellular carcinoma.

Abstract

AIM:

To investigate the effects of Axl deglycosylation on tumor lymphatic metastases in mouse hepatocellular carcinoma cell lines.

METHODS:

Western blotting was used to analyze the expression profile of Axl glycoprotein in mouse hepatocellular carcinoma cell line Hca-F treated with tunicamycin and PNGase F 3-(4,5)-dimethylthiazol(-zyl)-3,5-diphenyltetrazolium bromide (MTT) assay, extracellular matrix (ECM) invasion assay (in vitro) and tumor metastasis assay (in vivo) were utilized to evaluate the effect of Axl deglycosylation on the Hca-F cell proliferation, invasion and lymphatic metastasis.

RESULTS:

Tunicamycin and PNGase F treatment markedly inhibited Axl glycoprotein synthesis and expression, proliferation, invasion, and lymphatic metastasis both in vitro and in vivo. In the MTT assay, proliferation was apparent in untreated Hca-F cells compared with treated Hca-F cells. In the ECM invasion assay (in vitro), treated cells passed through the ECMatrix gel in significantly smaller numbers than untreated cells (tunicamycin 5 μg/mL: 68 ± 8 vs 80 ± 9, P = 0.0222; 10 μg/mL: 50 ± 6 vs 80 ± 9, P = 0.0003; 20 μg/mL: 41 ± 4 vs 80 ± 9, P = 0.0001); (PNGase F 8 h: 66 ± 7 vs 82 ± 8, P = 0.0098; 16 h: 49 ± 4 vs 82 ± 8, P = 0.0001; 24 h: 34 ± 3 vs 82 ± 8, P = 0.0001). In the tumor metastasis assay (in vivo), average lymph node weights of the untreated Hca-F group compared with treated Hca-F groups (tunicamycin 5 μg/mL: 0.84 ± 0.21 g vs 0.72 ± 0.19 g, P = 0.3237; 10 μg/mL: 0.84 ± 0.21 g vs 0.54 ± 0.11 g, P = 0.0113; 20 μg/mL: 0.84 ± 0.21 g vs 0.42 ± 0.06 g, P = 0.0008); (PNGase F 8 h: 0.79 ± 0.15 g vs 0.63 ± 0.13 g, P = 0.0766; 16 h: 0.79 ± 0.15 g vs 0.49 ± 0.10 g, P = 0.0022; 24 h: 0.79 ± 0.15 g vs 0.39 ± 0.05 g, P = 0.0001). Also, average lymph node volumes of the untreated Hca-F group compared with treated Hca-F groups (tunicamycin 5 μg/mL: 815 ± 61 mm³ vs 680 ± 59 mm³, P = 0.0613; 10 μg/mL: 815 ± 61 mm³ vs 580 ± 29 mm³, P = 0.0001; 20 μg/mL: 815 ± 61 mm³ vs 395 ± 12 mm³, P = 0.0001); (PNGase F 8 h: 670 ± 56 mm³ vs 581 ± 48 mm³, P = 0.0532; 16 h: 670 ± 56 mm³ vs 412 ± 22 mm³, P = 0.0001; 24 h: 670 ± 56 mm³ vs 323 ± 11 mm³, P = 0.0001).

CONCLUSION:

Alteration of Axl glycosylation can attenuate neoplastic lymphatic metastasis. Axl N-glycans may be a universal target for chemotherapy.

• Authors: Ji Li, Li Jia, Zhen-Hai Ma, Qiu-Hong Ma, Xiao-Hong Yang, Yong-Fu Zhao
• Journal: World journal of gastroenterology (World J Gastroenterol) Vol. 18 Issue 38 Pg. 5369-76 (Oct 14 2012) ISSN: 2219-2840 [Electronic] United States
• PMID: 23082053 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Biomarkers, Tumor
• Proto-Oncogene Proteins
• Receptor Protein-Tyrosine Kinases
• Axl Receptor Tyrosine Kinase
• AXL receptor tyrosine kinase, mouse

Topics

• Animals
• Biomarkers, Tumor (metabolism)
• Blotting, Western
• Carcinoma, Hepatocellular (metabolism, pathology)
• Cell Line, Tumor
• Cell Proliferation
• Glycosylation
• Liver Neoplasms (metabolism, pathology)
• Liver Neoplasms, Experimental (metabolism, pathology)
• Lymphatic Metastasis
• Mice
• Neoplasm Invasiveness
• Proto-Oncogene Proteins (metabolism)
• Receptor Protein-Tyrosine Kinases (metabolism)
• Axl Receptor Tyrosine Kinase