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RelB plays an oncogenic role and conveys chemo-resistance to DLD-1 colon cancer cells
Xiaojun Zhou1, Zhili Shan1, Hengying Yang1
11Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, 215006 China.
Background:
Nuclear transcription factor kappa B (NF-κB) subunits exhibit crucial roles in tumorigenesis and chemo-sensitivity. Recent studies suggest that RelB, the key subunit of the alternative NF-κB pathway, plays a critical role in the progression of diverse human malignancies. However, the significance of RelB in colorectal cancer (CRC) remains unclear. Here, we systematically explored the functions of the alternative NF-κB subunit RelB in colon cancer cells and its underlying mechanism.
Methods:
Stably transfected RelB-shRNA DLD-1 cells were established using Lipofectamine 2000. NF-κB DNA-binding capability was quantified using an ELISA-based NF-κB activity assay. Cell growth was monitored by an x-Celligence system. Cell proliferation was analyzed by a CCK-8 and a Brdu proliferation assay. Response to 5-FU was assessed by an x-Celligence system. Cell apoptosis and cell cycle was detected using flow cytometry analyses. Cell migration and invasion abilities were detected by an x-Celligence system, Transwell inserts, and wound-healing assays. RelB expression and its clinical significance were analyzed using the CRC tissue microarray. The expression of NF-κB signaling subunits, AKT/mTOR signaling molecules, cell cycle related proteins, MMP2, MMP9, and Integrin β-1 were measured by Western blotting analyses.
Results:
The RelB-silencing inhibited cell growth of DLD-1 cells. The RelB-silencing exerted the anti-proliferative by downregulation of AKT/mTOR signaling. The RelB-silencing caused G0-G1 cell cycle arrested likely due to decreasing the expression of Cyclin D1 and CDK4, concomitant with increased expression of p27Kip1. The RelB-silencing enhanced cytotoxic effect of 5-FU and induced cell accumulation in S-phase. The RelB-silencing impaired the migration and invasion potential of DLD-1 cells, which was related to downregulation of MMP2, MMP9, and Integrin β-1. Importantly, the RelB expression was correlated with depth of tumor invasion, lymph node metastasis, metastasis stage, and pTNM stage. High-RelB expression was significantly correlated with poor overall survival in CRC patients.
Conclusion:
Our studies here provided evidence that RelB plays an oncogenic role and conveys chemo-resistance to 5-FU. RelB can be considered as an independent indicator of prognosis in CRC.
Insights
Nuclear transcription factor kappa B (NF-κB) subunit RelB promotes colorectal cancer (CRC) growth and chemo-resistance. Silencing RelB inhibits CRC progression and enhances 5-FU efficacy, suggesting RelB as a poor prognostic indicator.
Area of Science:
- Molecular Biology
- Oncology
- Cancer Research
Background:
- Nuclear transcription factor kappa B (NF-κB) subunits are implicated in tumorigenesis and chemo-sensitivity.
- RelB, a key subunit of the alternative NF-κB pathway, is crucial in various human malignancies, but its role in colorectal cancer (CRC) is not fully understood.
Purpose of the Study:
- To systematically investigate the functions of the alternative NF-κB subunit RelB in colon cancer cells.
- To elucidate the underlying mechanisms by which RelB influences colorectal cancer progression and chemo-resistance.
Main Methods:
- Established RelB-silencing in DLD-1 colon cancer cells using shRNA.
- Assessed NF-κB activity, cell growth, proliferation, apoptosis, cell cycle, migration, and invasion.
- Analyzed RelB expression in CRC tissues and correlated it with clinical parameters and patient survival.
Main Results:
- RelB silencing inhibited cell growth, proliferation, migration, and invasion, linked to AKT/mTOR pathway downregulation and reduced MMP2/MMP9/Integrin β-1 expression.
- RelB silencing induced G0-G1 cell cycle arrest and enhanced the cytotoxic effect of 5-fluorouracil (5-FU).
- High RelB expression correlated with advanced tumor invasion, lymph node metastasis, and poor overall survival in CRC patients.
Conclusions:
- RelB plays a significant oncogenic role in colorectal cancer, promoting tumor progression and conferring resistance to 5-FU chemotherapy.
- RelB can be considered an independent prognostic indicator for colorectal cancer patients.
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