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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DOI：
10.3748/wjg.v18.i38.5369
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发表时间：
2012-10-14
影响因子：
4.3
通讯作者：
Zhao, Yong-Fu
中科院分区：
文献类型：
--
作者：
Li, Ji;Jia, Li;Zhao, Yong-Fu
目的：探讨Axl去糖基化对小鼠肝癌细胞淋巴道转移的影响。用Western blotting和PNGase F3-（4，5）-dimethylthiazol（-zyl）-3，5-diphenyltetrazolium bromide（MU）法分析衣霉素处理后小鼠肝癌细胞系Hca-F中Axl糖蛋白的表达谱。细胞外基质（ECM）侵袭测定体外实验和体内肿瘤转移实验观察Axl去糖基化对Hca-F细胞增殖、侵袭和淋巴结转移的影响。衣霉素和PNGase F处理显著抑制Axl糖蛋白的合成和表达、增殖、侵袭和淋巴转移。在MTT试验中，与处理的Hca-F细胞相比，未处理的Hca-F细胞的增殖是明显的。在ECM侵袭测定中，（体外），经处理的细胞通过ECMatrix凝胶的数量显著少于未经处理的细胞（衣霉素5 μ g/mL：68 +/-8 vs 80 +/-9，P = 0.0222; 10 μ g/mL：50 +/-6 vs 80 +/-9，P = 0.0003; 20 μ g/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）。在肿瘤转移测定（体内）中，未处理的Hca-F组的平均淋巴结重量与处理的Hca-F组相比，（衣霉素5 μ g/mL：0.84 +/-0.21 g vs 0.72 +/-0.19 g，P = 0.3237; 10 μ g/mL：0.84 +/-0.21 g vs 0.54 +/-0.11 g，P = 0.0113; 20 μ g/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）。此外，未治疗的Hca-F组与治疗的Hca-F组相比，（衣霉素5 μ g/mL：815 +/-61 mm（3）vs 680 +/-59 mm（3），P = 0.0613; 10 μ g/mL：815 +/-61 mm（3）vs 580 +/-29 mm（3），P = 0.0001; 20 μ g/mL：815 ± 61 mm（3）vs 395 ± 12 mm（3），P = 0.0001）;（PNGase F 8 h：670 +/-56 mm（3）vs 581 +/-48 mm（3），P = 0.0532; 16 h：670 +/-56 mm（3）vs 412 +/-22 mm（3），P = 0.0001; 24 h：结论：Axl糖基化改变可抑制肿瘤淋巴结转移。Axl N-聚糖可能是化疗的通用靶标。
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 (MU) 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 mu g/mL: 68 +/- 8 vs 80 +/- 9, P = 0.0222; 10 mu g/mL: 50 +/- 6 vs 80 +/- 9, P = 0.0003; 20 mu g/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 mu g/mL: 0.84 +/- 0.21 g vs 0.72 +/- 0.19 g, P = 0.3237; 10 mu g/mL: 0.84 +/- 0.21 g vs 0.54 +/- 0.11 g, P = 0.0113; 20 mu g/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 mu g/mL: 815 +/- 61 mm(3) vs 680 +/- 59 mm(3), P = 0.0613; 10 mu g/mL: 815 +/- 61 mm(3) vs 580 +/- 29 mm(3), P = 0.0001; 20 mu g/mL: 815 +/- 61 mm(3) vs 395 +/- 12 mm(3), P = 0.0001); (PNGase F 8 h: 670 +/- 56 mm(3) vs 581 +/- 48 mm(3), P = 0.0532; 16 h: 670 +/- 56 mm(3) vs 412 +/- 22 mm(3), P = 0.0001; 24 h: 670 +/- 56 mm(3) vs 323 +/- 11 mm(3), P = 0.0001).CONCLUSION: Alteration of Axl glycosylation can attenuate neoplastic lymphatic metastasis. Axl N-glycans may be a universal target for chemotherapy.