A novel dynamic matrix detachment model reveals a shift from apoptosis to necrosis in melanoma cells.
Anchorage-independence is a hallmark of invasive cancer. The setback of the classical poly-HEMA static matrix detachment (SMD) anoikis model is the absence of dynamic fluid circulation, resulting in cell aggregates. We addressed this problem by developing a novel 3D cell culture dynamic matrix detachment (DMD) model with a turbulent-free laminar flow, yielding a very low shear stress. In this study, we focused on melanoma cells where apoptosis was evaluated both via annexin V flow cytometry and caspase cleavage. The DMD model was superior to SMD in the induction of melanoma cell death and in revealing a shift from apoptosis to necrotic cell death, as evident by failure to activate caspase 9 and a decrease in annexin V stain. Combination of DMD with cisplatin could further accentuate necrotic cell death in cisplatin-resistant melanoma cells. Thus, the DMD model may be a useful matrix deprivation model to identify necrotic vs. apoptotic cell death pathways.
Tzukert
Keren
K
Department of Medicine, Hadassah-Hebrew University Medical Center, P.O. Box 12000, 91120 Jerusalem, Israel.
Gorodestky
Raphael
R
Avrahami
Idit
I
Krasny
Lina
L
Shimony
Nilly
N
Elkin
Gregory
G
Nettelbeck
Dirk M
DM
Haviv
Yosef S
YS
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D008546:Melanoma, Experimental / Q000473:pathology*; D051379:Mice; D008954:Models, Biological; D009336:Necrosis