Tumour cell-derived WNT5B modulates in vitro lymphangiogenesis via induction of partial endothelial-mesenchymal transition of lymphatic endothelial cells.
Metastasis of the cervical lymph nodes frequently leads to poor survival of patients with oral squamous cell carcinoma (OSCC). The underlying mechanisms of lymph node metastasis are unclear. Wingless-type MMTV integration site family, member 5B (WNT5B), one component of the WNT signal pathway, was markedly up-regulated in OSCC sublines with high potential of lymphatic metastasis compared to that in OSCC cells with low nodal metastasis. Increased WNT5B mRNA was demonstrated in human OSCC tissues in comparison with adjacent non-tumorous tissues. Interestingly, the high level of WNT5B protein in serum was associated with lymph node metastasis in OSCC patients. Knockdown of WNT5B expression in OSCC sublines did not affect tumour growth but impaired lymph node metastasis and tumour lymphangiogenesis of orthotopic transplantation. Conditioned medium from WNT5B knockdown cells reduced the tube formation of lymphatic endothelial cells (LECs). In contrast, recombinant WNT5B enhanced the tube formation, permeability and migration of LECs. In LECs stained with phalloidin, the morphology of those treated with recombinant WNT5B changed from flat to spindle-like. Recombinant WNT5B also increased α-smooth muscle actin and inhibited the expression of vascular endothelial-cadherin but retained characteristics of endothelial cells. The results suggest that WNT5B functions in the partial endothelial-mesenchymal transition (EndoMT). Furthermore, WNT5B-induced tube formation was impaired in the LECs following the knockdown of EndoMT-related transcription factor, SNAIL or SLUG. The WNT5B-induced expression of Snail or Slug was abolished by IWR-1-endo and Rac1 inhibitors, which are involved in the WNT/β-catenin and planar cell polarity pathways, respectively. Collectively, the data suggest that WNT5B induces tube formation by regulating the expression of Snail and Slug proteins through activation of canonical and non-canonical WNT signalling pathways.
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
S-H
SH
Wang
National Institute of Cancer Research, National Health Research Institutes, Tainan, Taiwan.
J S
JS
Chang
Department of Otolaryngology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
J-R
JR
Hsiao
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
Y-C
YC
Yen
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
S S
SS
Jiang
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
S-H
SH
Liu
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
Y-L
YL
Chen
Pathology Core Laboratory, National Health Research Institutes, Miaoli, Taiwan.
Y-Y
YY
Shen
Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
J-Y
JY
Chang
National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.|PhD Program for Aging, Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan.
Y-W
YW
Chen
D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D042583:Lymphangiogenesis / Q000235:genetics; D008207:Lymphatic Metastasis; D008297:Male; D051379:Mice; D009062:Mouth Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D015398:Signal Transduction; D000071250:Snail Family Transcription Factors / Q000378:metabolism; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*; D060449:Wnt Signaling Pathway; D023041:Xenograft Model Antitumor Assays
C000604407:SNAI1 protein, human; D000071250:Snail Family Transcription Factors; C435831:WNT5B protein, human; D051153:Wnt Proteins