Assaying Wnt5A-mediated invasion in melanoma cells.
Wnt5A has been implicated in melanoma metastasis, and the progression of other cancers including pancreatic, gastric, prostate, and lung cancers. Assays to test motility and invasion include both in vivo assays and in vitro assays. The in vivo assays include the use of tail vein or footpad injections of metastatic cells, and are often laborious and expensive. In vitro invasion assays provide quick readouts that can help to establish conditions that either activate or inhibit melanoma cell motility, and to assess whether the conditions in question are worth translating into an in vivo model. Here we describe two standard methods for assaying motility and invasion in vitro including wound healing assays and Matrigel invasion assays (Boyden chamber assays). In addition, we and several other laboratories have previously shown that melanoma cells require matrix metalloproteinase (MMP)-2 for their invasion, and have recently shown that Wnt5A treatment can increase the levels of this enzyme in melanoma cells, as demonstrated by gelatin zymography. The use of these techniques can help to assess the migratory capacity of melanoma cells in response to Wnt treatment.
O'Connell
Michael P
MP
Laboratory of Immunology, National Institutes of Health, National Institute on Aging, Gerontology Research Center, Baltimore, MD, USA.
French
Amanda D
AD
Leotlela
Poloko D
PD
Weeraratna
Ashani T
AT
D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D004338:Drug Combinations; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology*; D009361:Neoplasm Invasiveness; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051153:Wnt Proteins / Q000378:metabolism*; D000071818:Wnt-5a Protein
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C059485:matrigel; D003094:Collagen; D011493:Protein Kinase C