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. 2020 Dec 31;22(1):371.
doi: 10.3390/ijms22010371.

MMP1 and MMP11 Expression in Peripheral Blood Mononuclear Cells upon Their Interaction with Breast Cancer Cells and Fibroblasts

Noemi Eiro  1 , Sandra Cid  1 , Nuria Aguado  2 , María Fraile  1 , Nagore de Pablo  1 , Berta Fernández  3 , Francisco Domínguez  4 , Luis O González  5 , Francisco J Vizoso  1   6
Affiliations

MMP1 and MMP11 Expression in Peripheral Blood Mononuclear Cells upon Their Interaction with Breast Cancer Cells and Fibroblasts

Noemi Eiro et al. Int J Mol Sci. .

Abstract

Tumor-infiltrating immune cells phenotype is associated with tumor progression. However, little is known about the phenotype of the peripheral blood mononuclear cells (PBMC) from breast cancer patients. We investigated MMP1 and MMP11 expression in PBMC from breast cancer patients and we analyzed gene expression changes upon their interaction with cancer cells and cancer-associated fibroblasts (CAF). We measured the impact of PBMC on proinflammatory gene expression in breast cancer cells, normal fibroblast (NF), and CAF and the impact on proliferation and invasiveness capacity of breast cancer cells. Gene expression of MMP1 and MMP11 in PBMC from breast cancer patients (n = 54) and control (n = 28); expression of IL1A, IL6, IL17, IFNβ, and NFĸB in breast cancer cell lines (MCF-7 and MDA-MB-231); and, additionally, IL10 and MMP11 in CAF and NF were analyzed by qRT-PCR before and after co-culture. Our results show the existence of a subpopulation of breast cancer patients (25.9%) with very high levels of MMP11 gene expression in PBMC. Also, gene expression of MMP1 and MMP11 increases in PBMC after co-culture with breast cancer cell lines, NF or CAF. PBMC from healthy or breast cancer patients induce an increased proliferation rate on MCF-7 and an increased invasiveness capacity of MDA-MB-231. Finally, we show a differential expression profile of inflammatory genes in NF and CAF when co-cultured with control or breast cancer PBMC. We have observed that MMPs' expression in PBMC is regulated by the microenvironment, while the expression of inflammatory genes in NF or CAF is differentially regulated by PBMC. These findings confirm the importance of the crosstalk between stromal cells and suggest that PBMC would play a role in promoting aggressive tumor behavior.

Keywords: PBMC; breast cancer; inflammation and cancer; inflammatory cells; stroma.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
MMP1 and MMP11 gene expression in peripheral blood mononuclear cells (PBMC) from breast cancer patients. MMP1 (A) and MMP11 (B) gene expression in PBMC from breast cancer patients (BC-PBMC, n = 54) and healthy women (controls, C-PBMC, n = 28). The horizontal line marks an arbitrary threshold, corresponding to the highest MMP11 gene expression in C-PBMC, used as cut-off point. Significant difference was found between MMP11 gene expression in BC-PBMC from patients above the threshold and C-PBMC. (n.s.: not significant). Results of the qRT-PCR data were represented as ΔΔCT values.
Figure 2
Figure 2
MMP1 and MMP11 gene expression in PBMC after co-culture with breast cancer cell lines. Scheme of experimental design (A). MMP1 gene expression in PBMC from healthy women (controls, C-PBMC, n = 6) and in PBMC from breast cancer patients (BC-PBMC, n = 4) before and after co-culture with MCF-7 (B) and MDA-MB-231 (C) breast cancer cell lines. MMP11 gene expression in C-PBMC before and after co-culture with MCF-7 and MDA-MB-231 breast cancer cell lines (n = 6) (D,E) and in BC-PBMC after the same co-culture conditions (n = 4) (F,G). (n.s.: not significant). Results of the qRT-PCR data were represented as ΔΔCT values.
Figure 3
Figure 3
MMP1 and MMP11 gene expression in PBMC after co-culture with normal fibroblasts (NF) and cancer-associated fibroblasts (CAF). Scheme of experimental design (A). MMP1 gene expression in PBMC from healthy women (controls, C-PBMC) before and after co-culture with NF (n = 6) and CAF (n = 11) (B,C), and in PBMC from breast cancer patients (BC-PBMC) before and after co-culture with NF (n = 6) or CAF (n = 24) (D,E). Comparative of the MMP1 gene expression in C-PBMC and BC-PBMC after co-culture with NF (n = 6) and CAF (n = 11) (F,G). Comparative of the MMP11 gene expression in C-PBMC and BC-PBMC after co-culture with NF (n = 6) or CAF (n = 24) (H,I). Data represent the mean ± SD. (* p ≤ 0.05; n.s.: not significant). Results of the qRT-PCR data were represented as ΔΔCT values.
Figure 4
Figure 4
Inflammatory profile of breast cancer cell lines and fibroblasts after co-culture with PBMC. Scheme of experimental design (A). Gene expression of IL1A, IL6, IL10, IL17, IFNβ, NFĸB, and MMP11 in NF (n = 6) (B) and CAF (n = 6) (C) after co-culture with PBMC from healthy women (controls, C-PBMC). Gene expression of IL1A, IL6, IL10, IL17, IFNβ, NFĸB, and MMP11 in NF (n = 6) (D) and CAF (n = 24) (E) after co-culture with PBMC from breast cancer patients. Comparative of IL1A, IL6, IL10, IL17, IFNβ, NFĸB, and MMP11 gene expression in NF (F) and CAF (G) after co-culture with C-PBMC or BC-PBMC. Data represent the mean ± SD. (* p ≤ 0.05; ** p ≤ 0.01). Results of the qRT-PCR data were represented as fold expression.
Figure 5
Figure 5
Relative proliferative capacity of breast cancer cell lines MCF-7 (A) and MDA-MB-231 (B) after cultured in presence of cm-PBMC, both from C-PBMC (n = 9) and BC-PBMC (n = 6). Invasive capacity of breast cancer cell lines MCF-7 (C) and MDA-MB-231 (D) after co-culture with C-PBMC and BC-PBMC in Matrigel invasion chambers. Data represent the mean ± SD. (* p ≤ 0.05).
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