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Updated: Sep 6, 2025
Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model
Published on: March 20, 2020
Comparative analysis of transient receptor potential channel 5 opposite strand-induced gene expression patterns and
Jinghui Peng1,2, Shengbin Pei1,2, Yangyang Cui1
1Department of Breast Surgery, The First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Abstract:
In patients with triple-negative breast cancer (TNBC), high tumour mutation burden and aberrant oncogene expression profiles are some of the causes of poor prognosis. Therefore, it is necessary to identify aberrantly expressed oncogenes, since they have the potential to serve as therapeutic targets. Transient receptor potential channel 5 opposite strand (TRPC5OS) has been previously shown to function as a novel tumour inducer. However, the underlying mechanism of TRPC5OS function in TNBC remain to be elucidated. Therefore, in the present study TRPC5OS expression was first measured in tissue samples of patients with TNBC and a panel of breast cancer cell lines (ZR-75-1, MDA-MB-453, SK-BR-3, JIMT-1, BT474 and HCC1937) by using qRT-PCR and Western blotting. Subsequently, the possible effects of TRPC5OS on MDA-MB-231 cells proliferation were determined using Cell Counting Kit-8 and 5-Ethynyl-2'-deoxyuridine assays after Lentiviral transfection of MDA-MB-231. In addition, potential interaction partners of TRPC5OS were explored using liquid chromatography-mass spectrometry (LC-MS)/MS. Gene expression patterns following TRPC5OS overexpression were also detected in MDA-MB-231 cells by using High-throughput sequencing. Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) analysis were then used to systematically verify the potential interactions among the TRPC5OS-regulated genes. The potential relationship between TRPC5OS-interacting proteins and gene expression patterns were studied using Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis. TRPC5OS expression was found to be significantly higher in TNBC tumour tissues and breast cancer cell lines compared with luminal tumour tissues and ZR-75-1. In addition, the overexpression of TRPC5OS significantly increased cell proliferation. High-throughput sequencing results revealed that 5,256 genes exhibited differential expression following TRPC5OS overexpression, including 3,269 upregulated genes and 1,987 downregulated genes. GO analysis results indicated that the functions of these differentially expressed genes were enriched in the categories of 'cell division' and 'cell proliferation' regulation. KEGG analysis showed that the TRPC5OS-regulated genes were associated with processes of 'homologous recombination' and 'TNF signalling pathways'. Subsequently, 17 TRPC5OS-interacting proteins were found using LC-MS/MS and STRING analysis. The most important protein among interacting proteins was ENO1 which was associated with glycolysis and regulated proliferation of cancer. In summary, data from the present study suggest that TRPC5OS overexpression can increase TNBC cell proliferation and ENO1 may be a potential target protein mediated by TRPC5OS. Therefore, TRPC5OS may serve as a novel therapeutic target for TNBC.
Insights
High expression of TRPC5OS drives triple-negative breast cancer (TNBC) proliferation. Targeting TRPC5OS and its interaction with ENO1 offers a potential therapeutic strategy for TNBC patients.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Triple-negative breast cancer (TNBC) is associated with poor prognosis due to factors like high tumor mutation burden and aberrant oncogene expression.
- Identifying novel therapeutic targets is crucial for improving outcomes in TNBC.
- Transient Receptor Potential Channel 5 Opposite Strand (TRPC5OS) has emerged as a potential tumor inducer, but its specific role in TNBC requires elucidation.
Purpose of the Study:
- To investigate the expression levels of TRPC5OS in TNBC.
- To explore the functional impact of TRPC5OS on TNBC cell proliferation.
- To identify TRPC5OS interaction partners and downstream molecular pathways involved in TNBC progression.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) and Western blotting were used to measure TRPC5OS expression in TNBC tissues and cell lines.
- Lentiviral transfection was employed to overexpress TRPC5OS in MDA-MB-231 cells, followed by proliferation assays (Cell Counting Kit-8, 5-Ethynyl-2'-deoxyuridine).
- Liquid chromatography-mass spectrometry (LC-MS/MS), high-throughput sequencing, Gene Ontology (GO), Kyoto Encyclopaedia of Genes and Genomes (KEGG), and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analyses were utilized to identify interaction partners and signaling pathways.
Main Results:
- TRPC5OS expression was significantly elevated in TNBC tissues and cell lines compared to luminal breast cancer tissues.
- Overexpression of TRPC5OS led to a significant increase in MDA-MB-231 cell proliferation.
- High-throughput sequencing identified 5,256 differentially expressed genes, with GO and KEGG analyses linking them to cell division, proliferation, homologous recombination, and TNF signaling pathways.
- LC-MS/MS and STRING analyses identified 17 TRPC5OS-interacting proteins, notably ENO1, which is implicated in glycolysis and cancer cell proliferation.
Conclusions:
- TRPC5OS overexpression promotes TNBC cell proliferation.
- ENO1 is identified as a key interacting protein potentially mediating TRPC5OS's oncogenic effects.
- TRPC5OS represents a promising novel therapeutic target for triple-negative breast cancer.
