Carcinogenic Parasite Secretes Growth Factor That Accelerates Wound Healing and Potentially Promotes Neoplasia
Fig 2
Ov-GRN-1 stimulated wound repair in vitro.
(A) Cholangiocytes exposed to ES products from flukes where Ov-grn-1 had been silenced by RNA interference displayed significantly reduced proliferation over 36 h of co-culture. ES products (10 μg/ml) were derived from flukes that were exposed to dsRNAs for 5 days. Cell proliferation was monitored in real time using xCELLigence; every tenth data point is shown to aid visualization. Statistical comparisons were between Ov-grn-1- and luc-dsRNA-treated parasites. (B) Images of the scratch assay involving H69 cholangiocyte monolayers co-cultured in Transwell plates with Ov-grn-1 or luc-dsRNA-treated. Dotted lines denote wound edges over time. (C) Selected time points were measured from the photographs in (B); statistical comparisons were between cells cultured with Ov-grn-1-and luc-dsRNAs. (D) Wound healing scratch assay as shown in panel c but using the CCA cell line M214 (D). (E) Wound healing scratch assay as shown in panels (C) and (D) but recombinant protein applied to cells instead of co-culturing cells with live flukes. Statistical comparisons were between 20 nM rTRX and rOv-GRN-1 treatments or rTRX and PBS treatments. For all panels, data points represent the averages of two or three biological replicates with 3–5 biological replicates displayed with SEM error bars (some bars masked by data points). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, ns = not significant. Additional data shown in S2 Fig.