Related Experiment Video
Updated: Feb 12, 2026
Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model
Published on: March 20, 2020
Effect of siRNA-mediated LGR5 Inhibition on the Expression of Stemness-related Genes in Triple-negative Breast Cancer
Sepideh Ghobakhloo1,2, Sima Jafarpour1,2, Reza Nedaeinia2
1Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Background:
Approximately 10%-15% of all breast cancer diagnoses are identified as triple-negative breast cancer (TNBC), a subtype known for its poor prognosis due to chemoresistance. TNBC lacks any receptors or proteins that are overexpressed, thus lacking targeted therapy. The protein LGR5, a marker for cancer stem cells and a promoter of Wnt signaling pathway activity, is notably upregulated in TNBC cases. LGR5 plays an important role in chemoresistance and tumorigenesis. Considering this, our research aim was to investigate the effect of LGR5 suppression by using specific siRNA in multicellular spheroid of the TNBC cell line and evaluation of stemness-related gene expression changes.
Materials And Methods:
Multicellular spheroids of the MDA-MB-231 TNBC cell line were prepared. The exosome was extracted from the human adipose mesenchymal stem cells (ADMSCs) and confirmed. Multicellular spheroids were separately transfected with siRNA, EXO-siRNA and, treated with cisplatin alone. Gene expression was studied using qRT-PCR.
Results:
Our findings revealed that inhibiting LGR5 with siRNA significantly reduces LGR5 expression. Furthermore, the results indicated a notable increase in the expression of stemness markers (SOX9, OCT4, and NANOG) in samples treated with cisplatin. However, the reduction of LGR5 expression via siRNA led to a marked decrease in SOX9 and OCT4 levels, while NANOG expression remained largely unchanged.
Conclusions:
In summary, the results indicated that LGR5 suppression is effective in reducing stemness-related genes and may be considered a good candidate for combination therapy along with chemotherapy.
Insights
Suppressing LGR5 in triple-negative breast cancer (TNBC) cells reduces stemness markers. LGR5 inhibition shows promise as a combination therapy to improve chemotherapy effectiveness for TNBC.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Triple-negative breast cancer (TNBC) is an aggressive subtype lacking targeted therapies due to absent overexpressed receptors.
- LGR5, a marker for cancer stem cells and Wnt pathway promoter, is upregulated in TNBC and linked to chemoresistance and tumorigenesis.
Purpose of the Study:
- To investigate the impact of LGR5 suppression using siRNA on stemness-related gene expression in TNBC multicellular spheroids.
- To evaluate LGR5's role in TNBC chemoresistance and tumorigenesis.
Main Methods:
- TNBC MDA-MB-231 cell line multicellular spheroids were cultured.
- Cells were treated with siRNA targeting LGR5, exosomes with siRNA (EXO-siRNA), or cisplatin.
- Gene expression of LGR5 and stemness markers (SOX9, OCT4, NANOG) was analyzed using qRT-PCR.
Main Results:
- siRNA-mediated LGR5 suppression significantly reduced LGR5 expression.
- Cisplatin treatment increased stemness markers SOX9, OCT4, and NANOG.
- LGR5 suppression decreased SOX9 and OCT4 expression, while NANOG levels remained relatively unchanged.
Conclusions:
- LGR5 suppression is an effective strategy to reduce stemness-related gene expression in TNBC.
- Targeting LGR5 may enhance chemotherapy efficacy, suggesting its potential as an adjunct therapy for TNBC.
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