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Nrf2 pathway regulates multidrug-resistance-associated protein 1 in small cell lung cancer
1Department of Pathology, Shanghai Medical College, Fudan University, Shanghai, China.
Abstract:
Although multidrug-resistance-associated protein-1 (MRP1) is a major contributor to multi-drug resistance (MDR), the regulatory mechanism of Mrp1 still remains unclear. Nrf2 is a transcription factor that regulates cellular defense response through antioxidant response elements (AREs) in normal tissues. Recently, Nrf2 has emerged as an important contributor to chemo-resistance in tumor tissues. In the present study, the role of Nrf2-ARE pathway on regulation of Mrp1 was investigated. Compared with H69 lung cancer cells, H69AR cells with MDR showed significantly higher Nrf2-ARE pathway activity and expression of Mrp1 as well. When Nrf2 was knocked down in H69AR cells, MRP1's expression decreased accordingly. Moreover, those H69AR cells with reduced Nrf2 level restored sensitivity to chemo-drugs. To explore how Nrf2-ARE pathway regulates Mrp1, the promoter of Mrp1 gene was searched, and two putative AREs--ARE1 and ARE2--were found. Using reporter gene and ChIP assay, both ARE1 and ARE2 showed response to and interaction with Nrf2. In 40 cases of cancer tissues, the expression of Nrf2 and MRP1 was measured by immunohistochemistry (IHC). As the quantitive data of IHC indicated, both Nrf2 and MRP1 showed significantly higher expression in tumor tissue than adjacent non-tumor tissue. And more important, the correlation analysis of the two genes proved that their expression was correlative. Taken together, theses data suggested that Nrf2-ARE pathway is required for the regulatory expression of Mrp1 and implicated Nrf2 as a new therapeutic target for MDR.
Insights
The Nrf2-ARE pathway regulates multidrug-resistance-associated protein-1 (MRP1) expression, a key factor in multi-drug resistance (MDR). Targeting Nrf2 may offer a new strategy to overcome MDR in cancer treatment.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Background:
- Multidrug-resistance-associated protein-1 (MRP1) is crucial for multi-drug resistance (MDR) in cancer.
- The regulatory mechanisms of MRP1, particularly in tumor tissues, remain incompletely understood.
- Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor involved in cellular defense and chemo-resistance.
Purpose of the Study:
- To investigate the role of the Nrf2-antioxidant response element (ARE) pathway in regulating MRP1 expression.
- To determine if Nrf2 influences MRP1 expression and MDR in lung cancer cells.
- To explore the therapeutic potential of targeting the Nrf2 pathway for overcoming MDR.
Main Methods:
- Comparison of Nrf2-ARE pathway activity and MRP1 expression in H69 and H69AR lung cancer cells.
- Nrf2 knockdown experiments in H69AR cells to assess effects on MRP1 expression and drug sensitivity.
- Analysis of the MRP1 gene promoter for putative AREs using reporter gene and ChIP assays.
- Immunohistochemical analysis of Nrf2 and MRP1 expression in human cancer tissues.
Main Results:
- H69AR cells with MDR exhibited higher Nrf2-ARE pathway activity and MRP1 expression than H69 cells.
- Nrf2 knockdown in H69AR cells led to decreased MRP1 expression and restored sensitivity to chemotherapy drugs.
- The MRP1 promoter contains functional AREs (ARE1 and ARE2) that interact with Nrf2.
- Both Nrf2 and MRP1 were significantly upregulated in tumor tissues compared to adjacent non-tumor tissues, with a positive correlation between their expression.
Conclusions:
- The Nrf2-ARE pathway is essential for the regulation of MRP1 expression.
- Nrf2 plays a significant role in mediating MRP1-dependent multi-drug resistance in cancer.
- Nrf2 represents a promising therapeutic target for overcoming chemo-resistance in MDR cancers.
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