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Following forced differentiation in culture the limbal epithelial cells revealed an increase in K5/14 gene/protein expression levels in concert with a predictable rise in a known differentiation marker.
Our data showed that forced differentiation of HPV-positive KCs resulted in the expression of TLR9, however, as HPVs inhibits differentiation this may appear as TLR9 loss similar to what was seen previously [12].
Furthermore K14/K5 expression levels were not lowered (in fact they increased) within a limbal epithelial cell culture undergoing forced differentiation suggesting K14/K5 is an unreliable maker for undifferentiated cells ex vivo.
Our hypothesis that Elf5 regulates the switch from mammary epithelial cell proliferation to differentiation is supported by the report of a mouse in which ectopic over-expression of Elf5 in the mammary gland led to a reduction in epithelial proliferation and forced differentiation of the mammary epithelium [19].
ACSVL3 expression was diminished by ∼80% following forced differentiation.
Forced differentiation was also used to sensitize GSC to radiotherapy.
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Forcing differentiation of CSCs may represent a potential effective therapeutic strategy.
Inhibition of the Notch signalling pathway using γ-secretase inhibitors can force differentiation of intestinal tumours induced by Apc loss into goblet cells.
Khan et al. recently reported that forcing differentiation can cause elimination of neoplastic clones (Khan et al., 2013); by contrast, our experiments show that restoring expression of the wing-fate regulator Vg suppresses tumorous overgrowth without inducing cell death.
To further confirm that the differentiation block was caused by Smg6 deletion, we forced EB differentiation via DMSO, which triggers fibroblastic or muscle-like cell differentiation, and retinoic acid (RA), which induced both ectoderm and endoderm differentiation in controls.
Overall, these observations demonstrated a potential oncorepressor activity of JMJD3 enzyme in vivo and in vitro in the setting of certain subtypes of AML such as M2 and M3, which was most likely in association with the forced myeloid differentiation as well as the reduced survival and proliferation of AML blasts.
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