Progression of Luminal Breast Tumors Is Promoted by Ménage à Trois between the Inflammatory Cytokine TNFα and the Hormonal and Growth-Supporting Arms of the Tumor Microenvironment

Table 1

Breast tumor cells that migrated out of tumor-spheroids following TNF𝛼 + Estrogen + EGF stimulation express reduced levels of E-cadherin.

E-cadherin expression	Cells dissociated mechanically from tumor-spheroids formed by non-stimulated cells	Cells dissociated mechanically from tumor-spheroids formed in the presence of TNF𝛼 + Estrogen + EGF stimulation	Single cells that have detached spontaneously from tumor-spheroids formed in the presence of TNF𝛼 + Estrogen + EGF stimulation
Mean fluorescence (MFI)	15.8	15.4	11.5
% Positive cells	18.0	20.9	5.7
Score (MFI × %)	284.4	321.9	65.5
Normalized values	1.00	1.13	0.23

The table summarizes the analyses performed for E-cadherin expression in tumor-spheroid assays. The formation of tumor-spheroids and the ability of TNF𝛼 + Estrogen + EGF-stimulated cells to migrate out of the tumor-spheroids were shown in Figure 9. Then, tumor-spheroids and single cells were separated by a 40 𝜇m nylon mesh. Cells from tumor-spheroids formed in the absence or in the presence of TNF𝛼 + Estrogen + EGF stimulation were dissociated mechanically by trypsinization and were stained for E-cadherin in comparison to cells that have detached spontaneously from the tumor-spheroids formed in the presence of TNF𝛼 + Estrogen + EGF stimulation. The three cell types were analyzed for surface expression of E-cadherin by flow cytometry analyses. The results are from a representative experiment of 𝑛≥3.