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.
Key wordsMelanoma Wnt5A Matrigel Boyden chamber Invasion Wound-healing assays
We thank Dr. Michel Bernier and Dr. Paritosh Ghosh for helpful comments on this manuscript. Any data represented in this chapter was generated with the support of funds from the Intramural Research Program of the National Institute on Aging, Baltimore, MD.
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