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Interleukin-12 Inhibits Tumor Growth and Metastasis Promoted by Tumor-Associated Mesenchymal Stem Cells in
Babak Jahangiri1, Zahra-Soheila Soheili1, Elahe Asadollahi2
1Department of Molecular Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Objective:
The aim of this study was to understand the interactions between tumor-associated mesenchymal stem cells (TA-MSCs) and triple-negative breast cancer (TNBC) cells, which appear to be necessary for developing effective therapies.
Materials And Methods:
In this experimental study, MDA-MB-231 and 4T1 TNBC cells were co-cultured with bone marrow-derived MSCs, and TA-MSCs conditioned media (CM) were collected. TA-MSC CM-treated TNBC cells were subjected to migration and invasion assays. Epithelial-mesenchymal transition (EMT) marker expression was quantified by real-time polymerase chain reaction (RT-PCR). Cell proliferation was measured using trypan blue exclusion technique, while cell cycle distribution and apoptosis were assessed by flow cytometry. The effects of TA-MSCs on tumor volume, survival rate, and lung metastasis were evaluated by subcutaneous co-injection of MSCs with 4T1 cells in the right flanks of BALB/c mice (n=5 per group). Intratumoral interleukin-12 (IL-12) immunotherapy was performed using lentiviral particles as a rescue experiment. The TA-MSCs RNA-seq dataset (PRJEB27694) was analyzed to detect elevated metastasis-associated oncogenes, downloaded from the European Nucleotide Archive database. For validation of the RNA-seq data analysis, the expression levels of candidate oncogenes were evaluated in TA-MSCs, TNBC cells, and tumor tissue using RT-PCR.
Results:
TA-MSCs enhanced migration, invasion, and EMT of TNBC cells in vitro without affecting cell proliferation or apoptosis. In vivo, TA-MSCs increased tumor growth and lung metastasis, while decreasing survival rates. IL-12 therapy elevated serum IL-12 and interferon-gamma (IFN-γ) expression, suppressed tumor volume and lung metastasis, and improved overall survival in the TA-MSC group. RNA-seq data analysis identified upregulated oncogenes in TA-MSCs, among which MMP3, CXCL2, CXCL5, and ICAM1 were selected as the most relevant to metastasis. These genes showed increased expression in TA-MSCs, TNBC cells, and tumor tissues.
Conclusion:
The findings of the present study revealed a complex interplay between TA-MSCs and TNBC cells that affects tumor growth and metastasis. Preclinical results indicate that intratumoral IL-12 immunotherapy shows promise in overcoming TA-MSC-promoted tumor growth and metastasis.
Insights
Tumor-associated mesenchymal stem cells (TA-MSCs) promote triple-negative breast cancer (TNBC) growth and metastasis. Intratumoral IL-12 immunotherapy effectively counteracts these effects, offering a promising therapeutic strategy for TNBC.
Area of Science:
- Oncology
- Cancer Biology
- Immunotherapy
Background:
- Triple-negative breast cancer (TNBC) is an aggressive subtype with limited treatment options.
- Tumor-associated mesenchymal stem cells (TA-MSCs) play a significant role in tumor progression.
- Understanding TA-MSC and TNBC cell interactions is crucial for developing novel therapies.
Purpose of the Study:
- To investigate the interaction between TA-MSCs and TNBC cells.
- To evaluate the impact of TA-MSCs on TNBC cell behavior in vitro and in vivo.
- To assess the therapeutic potential of IL-12 immunotherapy in overcoming TA-MSC-mediated tumor promotion.
Main Methods:
- Co-culture of TNBC cells (MDA-MB-231, 4T1) with bone marrow-derived MSCs.
- In vitro assays for migration, invasion, proliferation, cell cycle, and apoptosis.
- In vivo studies involving subcutaneous co-injection in mice to assess tumor growth, metastasis, and survival.
- RNA-sequencing (RNA-seq) analysis of TA-MSCs to identify metastasis-associated oncogenes.
- Intratumoral IL-12 immunotherapy as a rescue experiment.
Main Results:
- TA-MSCs enhanced TNBC cell migration, invasion, and epithelial-mesenchymal transition (EMT) in vitro.
- In vivo, TA-MSCs promoted tumor growth, lung metastasis, and reduced survival rates.
- IL-12 immunotherapy successfully suppressed tumor growth and metastasis while improving survival.
- Upregulated oncogenes (MMP3, CXCL2, CXCL5, ICAM1) in TA-MSCs were identified and validated.
Conclusions:
- TA-MSCs exhibit a complex interplay with TNBC cells, driving tumor growth and metastasis.
- Intratumoral IL-12 immunotherapy demonstrates significant preclinical efficacy against TA-MSC-promoted TNBC.
- Targeting TA-MSC-mediated pro-tumorigenic effects is a viable therapeutic strategy.
Related Concept Videos
01:19Mesenchymal Stem Cells
02:17The Tumor Microenvironment
02:30Metastasis
Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...