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Published on: March 6, 2018
Cyto-genotoxic assessment of bisphenol P through expression of DNA damage/repair genes in MDBK cell line
Muhammad Muddassir Ali1, Samra Afzal2, Maryam Javed2
1Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, 54000, Pakistan. muddassir.ali@uvas.edu.pk.
Abstract:
Bisphenol P (BPP) is a recognized endocrine disruptor with detrimental effects on human health. This study aimed to evaluate BPP's cytotoxic and genotoxic effects on Madin-Darby bovine kidney (MDBK) cells by examining changes in gene expression, genotoxicity, and cell survival. Various assays were employed, including the MTT assay, comet assay, micronucleus assay, and real-time PCR for gene expression analysis. Among the series of concentrations (0.5 µM, 1 µM, 2 µM, 4 µM, 8 µM, 16 µM, 32 µM, 64 µM, 128 µM, and 256 µM), the treatment with 32 µM BPP (LC50) resulted in 50% cell viability after 24 h via MTT assay. The comet assay revealed a significant increase in comet tail length in BPP-treated groups compared to controls, indicating DNA with the highest damage at the 3xLC50/2 dose concentration of BPP. The frequency of micronuclei (MNi) was higher than binuclei. A significantly higher level of cytokinesis-block proliferation index (CBPI) was also observed at higher doses than in the negative control group. Gene expression analysis indicated increased levels of OGG1 and HPRT1 in BPP-treated cells compared to untreated controls, with a dose-dependent elevation in OGG1 expression involved in DNA damage response. This study concluded that BPP exhibits both cytotoxic and genotoxic effects on MDBK cells. Expression of DNA repair genes (OGG1, HPRT1) served as biomarkers for genotoxicity. Furthermore, it is recommended that additional studies on BPP's molecular toxicity and its cross-species effects should be explored further to combat its harmful effects.
Insights
Bisphenol P (BPP) is a toxic endocrine disruptor. This study found BPP causes significant cell damage and DNA mutations in bovine kidney cells, highlighting its genotoxic risks.
Area of Science:
- Toxicology
- Molecular Biology
- Cell Biology
Background:
- Bisphenol P (BPP) is an endocrine disruptor with known adverse health effects.
- Understanding BPP's impact on cellular functions is crucial for risk assessment.
Purpose of the Study:
- To evaluate the cytotoxic and genotoxic effects of Bisphenol P (BPP) on Madin-Darby bovine kidney (MDBK) cells.
- To investigate BPP's impact on cell survival, DNA integrity, and gene expression.
Main Methods:
- Cytotoxicity assessed using MTT assay.
- Genotoxicity evaluated via comet assay and micronucleus assay.
- Gene expression analysis of DNA repair genes (OGG1, HPRT1) using real-time PCR.
Main Results:
- Bisphenol P (BPP) demonstrated cytotoxicity, with LC50 at 32 µM.
- Significant DNA damage observed in BPP-treated cells, indicated by increased comet tail length and micronuclei frequency.
- Elevated expression of DNA repair genes (OGG1, HPRT1) in response to BPP exposure.
Conclusions:
- Bisphenol P (BPP) exhibits significant cytotoxic and genotoxic effects on MDBK cells.
- OGG1 and HPRT1 gene expression can serve as biomarkers for BPP-induced genotoxicity.
- Further research into BPP's molecular toxicity and cross-species effects is warranted.
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