1. Recombinant OPCML protein effect in vitro.
Recombinant OPCML protein effect in vitro. A, schematic diagram (left) of OPCML and rOPCML, SDS–PAGE analysis of rOPCML expressed in Escherichia coli. Lane 1, Coomassie Blue; lane 2, rOPCML Western blot. B, rOPCML targets cancer but not normal cells (left); OSE-C2 and SKOV-3 cells were subjected to various concentrations of rOPCML (0.5, 1, 2, 5, and 10 μM). The MTT proliferation assay is shown relative to control vehicle-treated (dotted line) at 48 hours. A dose-dependent inhibition of SKOV-3 cell growth demonstrates the specificity of rOPCML for OPCML-deficient cells (NS, nonsignificant, *P = 0.004, **P < ). C, the OSE-C2 line and a panel of ovarian cancer lines (SKOV-3, IGROV, OVISE, OVCAR-5, A2780, PEA1, and PEA2) were exposed to 10 μM of rOPCML for 24 hours (white bar) and 48 hours (black bar) with MTT cell growth was normalized to vehicle-only controls (*P < , αP = , and βP = ). These data demonstrate that 6 of 7 (86%) of the ovarian cancer lines (but not OSE-C2) were significantly and profoundly growth suppressed after exposure to rOPCML. D, confocal coimmunofluorescence demonstrates the relative abundance of HER2 protein in SKOV-3 cells after either PBS application (top) or rOPCML application (bottom) with punctate colocalization of OPCML with HER2 in the latter.
Arthur B. McKie et al. Cancer Discovery 2012;2:
©2012 by American Association for Cancer Research