首页|Axl glycosylation mediates tumor cell proliferation,invasion and lymphatic metastasis in murine hepatocellular carcinoma
Axl glycosylation mediates tumor cell proliferation,invasion and lymphatic metastasis in murine hepatocellular carcinoma
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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; 10μg/mL:50 ± 6 vs 80 ± 9,P =0.0003; 20 lμg/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 ± 0.15g vs 0.49 ± 0.10 g,P =0.0022; 24 h:0.79 ± 0.15 g vs0.39 ± 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 ± 61mm3 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; 16h: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.
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