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Cisplatin-induced alterations in the expression of the mRNAs for UV-damage recognition protein
A Vaisman1, S Keeney, A F Nichols
1Department of Biochemistry & Biophysics, School of Medicine, University of North Carolina, Chapel Hill 27599-7260, USA.
Abstract:
Enhanced DNA repair is believed to be an important mechanism of the cisplatin-resistant phenotype. UV-damage recognition protein (UV-DRP) recognizes and binds to DNA lesions and may play a role in DNA nucleotide excision repair and/or replicative bypass (which is associated with post-replication repair). Potential alternations in the expression of mRNAs for UV-DRP were analyzed in this study. Two pairs of parental and cisplatin-resistant human ovarian carcinoma cell lines were utilized. Gene expression level was assessed by northern blot hybridization. No alterations in mRNA levels for the large subunit of UV-DRP were found following cisplatin treatment, whereas mRNA levels for the small subunit of UV-DRP were induced up to 4.5-fold. The time-course and concentration-response of this induction corresponded to the previously reported increase in the UV-DRP binding activity, as measured by gel shift assay. UV-DRP binding activity in cell extracts corresponds to expression of small subunit mRNA but not to expression of large subunit mRNA. These data suggest that the small subunit may be limiting for UV-DRP activity.
Insights
Cisplatin resistance in ovarian cancer may involve enhanced DNA repair. The study found that the small subunit of UV-damage recognition protein (UV-DRP) mRNA significantly increased in resistant cells, suggesting it limits UV-DRP activity.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- Cisplatin resistance is a major challenge in ovarian cancer treatment.
- Enhanced DNA repair mechanisms are implicated in cisplatin resistance.
- UV-damage recognition protein (UV-DRP) plays a role in DNA repair pathways.
Purpose of the Study:
- To investigate the role of UV-damage recognition protein (UV-DRP) expression in cisplatin resistance.
- To analyze alterations in mRNA levels of UV-DRP subunits in cisplatin-resistant ovarian cancer cells.
Main Methods:
- Utilized two pairs of parental and cisplatin-resistant human ovarian carcinoma cell lines.
- Assessed gene expression levels using northern blot hybridization.
- Measured UV-DRP binding activity via gel shift assay.
Main Results:
- No significant alterations in the large subunit of UV-DRP mRNA were observed.
- mRNA levels for the small subunit of UV-DRP were induced up to 4.5-fold in cisplatin-resistant cells.
- The induction of small subunit mRNA correlated with increased UV-DRP binding activity.
Conclusions:
- The small subunit of UV-DRP may be a limiting factor for UV-DRP activity in cisplatin-resistant ovarian cancer.
- Findings suggest a potential role for UV-DRP modulation in overcoming cisplatin resistance.