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Page 8 of 17 Mooney et al. J Cancer Metastasis Treat 2019;5:19 I http://dx.doi.org/10.20517/2394-4722.2018.93

A

B

C

Figure 3. Reversal of gene expression of metastatic MDA-MB-231 breast cancer cells (BCCs) after exposure to embryonic stem cell
(ESC)-microstrands at the transcriptional level as revealed by qRT-PCR analysis. A: After 72 h, significantly decreased TGF-α and
increased NKD2 gene expression in BCCs co-cultured with ESC-microstrands compared to both empty microstrand and non-co-cultured
controls; B: after 72 h, EGFR and vimentin gene expression decreased significantly compared to both controls; C: after 48 h, β-catenin and
GAPDH gene expression declined in BCCs co-cultured with ESC-microstrands compared to the both controls (****P < 0.0001; ***P < 0.001:
**P < 0.01; *P < 0.05)

decline in both EGFR and vimentin mRNA levels in the BCCs co-cultured with ESC-microstrands [Figure 3B].
There was also a decline in expression in the empty microstrand group for both EGFR and vimentin, but
it is not as extreme as for the ESC-microstrand group. Finally, both β-catenin and GAPDH mRNA levels
decreased in the BCCs after co-culture with ESC-microstrands for 48 h [Figure 3C]. This decrease persisted
for β-catenin, but not for GAPDH, after 72 h (data not shown).

ESC-microstrands reverse EGFR and β-catenin protein expression in metastatic BCCs
We further examined expression of EGFR, β-catenin, NKD2, and TGF-α in metastatic MDA-MB-231
BCCs at protein level after 48 h of co-culture with ESC-microstrands, in comparison with non-invasive