Partial Inhibition of Epithelial-to-Mesenchymal Transition (EMT) Phenotypes by Placenta-Derived DBMSCs in Human Breast Cancer Cell Lines, In Vitro

Abstract

Stem cell-based therapies hold significant potential for cancer treatment due to their unique properties, including migration toward tumor niche, secretion of bioactive molecules, and immunosuppression. Mesenchymal stem cells (MSCs) from adult tissues can inhibit tumor progression, angiogenesis, and apoptosis of cancer cells. We have previously reported the isolation and characterization of placenta-derived decidua basalis mesenchymal stem cells (DBMSCs), which demonstrated higher levels of pro-migratory and anti-apoptotic genes, indicating potential anti-cancer effects. In this study, we analyzed the anti-cancer effects of DBMSCs on human breast cancer cell lines MDA231 and MCF7, with MCF 10A used as control. We also investigated how these cancer cells lines affect the functional competence of DBMSCs. By co-culturing DBMSCs with cancer cells, we analyzed changes in functions of both cell types, as well as alterations in their genomic and proteomic profile. Our results showed that treatment with DBMSCs significantly reduced the functionality of MDA231 and MCF7 cells, while MCF 10A cells remained unaffected. DBMSC treatment decreased epithelial-to-mesenchymal transition (EMT)-related protein levels in MDA231 cells and modulated expression of other cancer-related genes in MDA231 and MCF7 cells. Although cancer cells reduced DBMSC proliferation, they increased their expression of anti-apoptotic genes. These findings suggest that DBMSCs can inhibit EMT-related proteins and reduce the invasive characteristics of MDA231 and MCF7 breast cancer cells, highlighting their potential as candidates for cell-based cancer therapies.

Keywords: EMT; MCF 10A; MCF7; MDA231; adhesion; decidua basalis MSCs; invasion; migration; placenta; proliferation.

Figure 1

Standardization of cellular ratio and co-culture time for treatment of DBMSCs with cancer cells: the effect of the cellular component of cancer cells at various ratios (1:1; 1:2, and 1:5) and at different time points (24 h, 48 h, and 96 h), with a measurable effect on DBMSCs, as recorded in MTS assay (A). Adhesion of DBMSCs decreased significantly after treatment with MCF7 and MDA231 cells for 96 h, but not with MCF 10A cells (B,B(i)). Proliferation of DBMSCs after co-culture with cancer cells at 1:5 ratio for 96 h decreased significantly with MCF7 and MDA231 cells (C,C(i)). Similarly, after treatment with the cellular component of MCF7 and MDA231 cells at a 1:5 ratio and incubation for 96 h, DBMSCs showed decreased migration (D,D(i)), as well as decreased invasion (E,E(i)), as recorded in the xCelligence assay, compared to MCF 10A and untreated control cells. The experiments were replicated three times with DBMSCs isolated from three different placentas. Bars represent standard errors. * p < 0.05.

Figure 2

Migration and invasion of DBMSCs after their co-culture with cancer cells: migration and invasion of DBMSCs after their treatment for 96 h with the cellular component of MCF 10A, MCF7, and MDA231 cells at a 1:5 ratio, further evaluated by Transwell assay. DBMSCs treated with MCF7 and MDA231 cells migrated at a significantly faster rate through the 8 µM pore of a Transwell filter compared to the MCF 10A cells and the untreated control. Panel (A) shows the photomicrograph (20X magnification) of the migrated cells under various treatment conditions, whereas the percentage of total migrated cells is presented in a bar graph (A(i)). Similar results were observed for the invasion assay performed over Matrigel-coated Transwell plates under similar treatment and incubation conditions (B,B(i)). DBMSCs isolated from three individual placentas were used in these experiments. The experiments were repeated three times for statistical analyses. Bars represent standard errors. * p < 0.05).

Figure 3

Effect of cancer cell treatment on the expression of chemokines and cytokines on DBMSCs: Flow cytometry analysis for expression of chemokines and cytokines by DBMSCs after co-culture with DBMSCs showed a significant increase in IL-21, IFNA2, TNSF13B, and IL-17R for 96 h (A) but a significant decrease in the expression level of IL-27. The mean fluorescence index (MFI) is plotted and shown as a bar diagram (B). Bars represent standard errors. * p < 0.05.

Figure 4

Standardization of co-culture time for treatment of cancer cells with DBMSCs: the effect of the cellular component of DBMSCs at different time points (24 h, 48 h, and 96 h) with a measurable effect on MCF 10A, MCF7, and MDA231 cells, as recorded in the MTS assay (A). Adhesion of all cancer cells as well as the control cells did not change after treatment with DBMSCs for 96 h (B,B(i)). Proliferation of MCF7 and MDA231 cells decreased significantly after co culture with DBMSCs at a 1:5 ratio for 96 h (C,C(i)). Similarly, after treatment with the cellular component of DBMSCs at a 1:5 ratio, and after incubation for 96 h, MCF7 and MD231, but not MCF 10A or the untreated controls, showed decreased migration (D,D(i)) as well as decreased invasion (E,E(i)), as recorded in the xCelligence assay. The experiments were replicated three times with DBMSCs isolated from three separate placentas. Bars represent standard errors. * p < 0.05.

Figure 5

Migration and invasion of cancer cells after their treatment with DBMSCs: MCF7 and MDA231 migration and invasion after their treatment with DBMSCs at a 1:5 ratio and for 96 h were further assessed by Transwell assays. MCF7 and MDA231 cells migrated at a significantly slower rate through the 8 µM pore of a Transwell membrane after their treatment with the cellular component of DBMSCs for 96 h at a 1:5 ratio, compared to the MCF 10A cells and untreated controls. Panel (A) shows the photomicrographs (20X magnification) of the migrated cancer cells, whereas the percentage of total migrated cells is presented in a bar graph (A(i)). Similar results were observed for an invasion assay performed over a Matrigel-coated Transwell membrane under similar treatment and incubation conditions (B,B(i)). DBMSCs isolated from three different placentas were used in these experiments. The experiments were repeated three times for statistical analyses. Bars represent standard errors. * p < 0.05).

Figure 6

Effect of DBMSC treatment on the expression of EMT effectors on MDA231 cells: Flow cytometry analysis for the expression of effectors responsible for EMT by MDA231 cells after co-culture with MDA231 cells for 96 h showed a significant increase in E-cadherin and WNT5A (A), and a significant decrease in the expression level of Snail, vimentin and fibronectin. The mean fluorescence index (MFI) is plotted and shown as a bar diagram (B). Bars represent standard errors. * p < 0.05.