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Published on: September 1, 2019
Runt-related transcription factor 1 promotes apoptosis and inhibits neuroblastoma progression in vitro and in vivo
1Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Background:
Runt-related transcription factor 1 (RUNX1) is a heterodimeric transcription factor that binds to the core element of many enhancers and promoters and can accelerate apoptosis in various tumors. However, the regulatory mechanisms underlying RUNX1 expression in neuroblastoma (NB), a highly malignant tumor in childhood, remain largely unclear. In this study, we aimed to assess the role of RUNX1 in NB and to reveal the underlying mechanisms that may contribute to finding a potential therapeutics strategy against NB.
Methods:
Growth, invasion, metastasis and angiogenesis were assessed using Cell Counting Kit-8 (CCK-8) immunocytochemistry, and studies involving soft agar, cell invasion, tube formation and whole animals. The levels of expression were measured using real-time quantitative PCR for RNA, Western blot and immunostaining analyses for proteins. Luciferase reporter and chromatin immunoprecipitation assays indicated that RUNX1 directly binds within the BIRC5, CSF2RB and NFKBIA promoter regions to facilitate transcription. The level of apoptosis was assessed by determining mitochondrial membrane potential and flow cytometry.
Results:
RUNX1 was highly expressed in ganglioneuroma (GN) and well-differentiated (WD) tissues relative to the poorly differentiated (PD) and undifferentiated (UD) ones. Moreover, RUNX1 effectively reduced cell viability, invasion, metastasis, angiogenesis, and promoted apoptosis in vitro and in vivo. RUNX1 reduced BIRC5 transcription and increased CSF2RB and NFKBIA transcription by directly binding BIRC5, CSF2RB and NFKBIA promoters. In addition, cytotoxic drugs, especially cisplatin, significantly increased RUNX1 expression in NB cells and promoted apoptosis.
Conclusions:
These data show that RUNX1 is an independent surrogate marker for the progression of NB and it can be used for monitoring NB prognosis during therapy.
Insights
Runt-related transcription factor 1 (RUNX1) acts as a tumor suppressor in neuroblastoma (NB). Upregulating RUNX1 inhibits NB progression and enhances apoptosis, suggesting its therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Runt-related transcription factor 1 (RUNX1) is a key transcription factor involved in various cellular processes, including apoptosis.
- The role and regulatory mechanisms of RUNX1 in neuroblastoma (NB), a prevalent childhood cancer, are not well understood.
- Investigating RUNX1 in NB is crucial for developing novel therapeutic strategies.
Purpose of the Study:
- To elucidate the function of RUNX1 in neuroblastoma.
- To identify the molecular mechanisms underlying RUNX1's role in NB progression.
- To explore RUNX1 as a potential therapeutic target for NB.
Main Methods:
- Assessed cell viability, invasion, metastasis, and angiogenesis using CCK-8, soft agar, invasion, and tube formation assays.
- Quantified gene and protein expression via real-time qPCR, Western blot, and immunostaining.
- Utilized luciferase reporter and chromatin immunoprecipitation assays to determine RUNX1's direct transcriptional regulation of target genes (BIRC5, CSF2RB, NFKBIA).
- Evaluated apoptosis using mitochondrial membrane potential and flow cytometry.
Main Results:
- RUNX1 expression was inversely correlated with NB differentiation, being higher in well-differentiated tissues.
- RUNX1 significantly suppressed NB cell viability, invasion, metastasis, and angiogenesis in vitro and in vivo.
- RUNX1 directly downregulated BIRC5 and upregulated CSF2RB and NFKBIA by binding to their promoter regions.
- Cisplatin treatment increased RUNX1 expression and promoted apoptosis in NB cells.
Conclusions:
- RUNX1 functions as an independent prognostic marker for neuroblastoma progression.
- RUNX1 plays a critical role in suppressing NB growth and metastasis.
- RUNX1 modulation, potentially enhanced by cytotoxic drugs like cisplatin, offers a promising therapeutic avenue for NB.
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