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[Expression of BRD7-interacting proteins,BRD2 and BRD3, in nasopharyngeal carcinoma tissues]
Ming Zhou1, Cong Peng, Xin-Min Nie
1Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410078, PR China.
Background & Objective:
BRD7 is a novel gene tightly associated with nasopharyngeal carcinoma(NPC) cloned by cDNA representational difference analysis (cDNA RDA). Two proteins,BRD2 and BRD3, including bromodomain and interacting with BRD7 protein had been screened from human fetal brain cDNA library by yeast two-hybrid system. This study was designed to further identify the interactions of BRD2 and BRD3 with BRD7 respectively and to investigate the expression and action pattern of BRD2 and BRD3 in NPC.
Methods:
BRD2 and BRD3 genes were respectively co-transformed to yeast Y187 with BRD7 gene, then the yeast cotransformers were blotted to nylon membrane. And then the expression of report gene Lac Z by beta-Gal was determined and the interactions of BRD2 and BRD3 proteins with BRD7 protein were identified. Besides,reverse transcription-polymerase chain reaction (RT- PCR) was used to examine the expression of BRD2 and BRD3 genes in normal nasopharyngeal epithelium and NPC biopsies, and to detect the effect of re-expression of BRD7 gene on the expression of BRD2 and BRD3 genes in HNE1 stably transfected BRD7 gene.
Results:
The yeast transformers showed all blue by yeast two-hybrid system, which further identified that BRD2 and BRD3 proteins could respectively interact with BRD7 protein. Down-expression or loss of BRD2 and BRD3 genes were detectable in NPC biopsies. The expression levels of BRD2 and BRD3 were increasing with the re-expression of BRD7 gene in HNE1 stably transfected with BRD7.
Conclusion:
BRD7 protein could respectively interact with proteins, BRD2 and BRD3, and BRD7 could up-regulate the expression levels of BRD2 and BRD3 genes in mRNA level to some extent. Each of these three homolog proteins might be capable of forming heteromers with the others, which play important roles in the suppression of growth of NPC cells.
Insights
Bromodomain-containing protein 7 (BRD7) interacts with BRD2 and BRD3, potentially forming complexes that suppress nasopharyngeal carcinoma (NPC) cell growth. BRD7 also up-regulates BRD2 and BRD3 expression in NPC.
Area of Science:
- Molecular biology
- Oncology
- Genetics
Background:
- Bromodomain-containing protein 7 (BRD7) is a novel gene associated with nasopharyngeal carcinoma (NPC).
- BRD2 and BRD3 proteins, interacting with BRD7, were identified using a yeast two-hybrid system.
- Understanding the interactions and expression patterns of BRD2, BRD3, and BRD7 is crucial for NPC research.
Purpose of the Study:
- To confirm the interactions between BRD7 and BRD2/BRD3 proteins.
- To investigate the expression patterns of BRD2 and BRD3 in NPC.
- To determine the effect of BRD7 on BRD2 and BRD3 expression in NPC cells.
Main Methods:
- Yeast two-hybrid system to identify protein interactions.
- Beta-galactosidase assay to confirm gene expression.
- Reverse transcription-polymerase chain reaction (RT-PCR) to analyze gene expression levels in normal and cancerous tissues.
- Stable transfection of HNE1 cells with BRD7 to study its effect on BRD2 and BRD3 expression.
Main Results:
- Yeast two-hybrid assays confirmed that BRD2 and BRD3 proteins interact with BRD7.
- Down-expression or loss of BRD2 and BRD3 genes was observed in NPC biopsies.
- Re-expression of BRD7 in HNE1 cells led to increased expression of BRD2 and BRD3 genes.
Conclusions:
- BRD7 protein interacts with both BRD2 and BRD3 proteins.
- BRD7 up-regulates the mRNA expression levels of BRD2 and BRD3.
- These three proteins may form heteromers that play a significant role in suppressing NPC cell growth.
