Genotoxic Effects of Cylindrospermopsin, Microcystin-LR and Their Binary Mixture in Human Hepatocellular Carcinoma (HepG2) Cell Line
Abstract
:1. Introduction
2. Results
2.1. Viability of HepG2 Cells Exposed to CYN, MCLR and Their Mixtures
2.2. The Formation of DNA Double Strand Breaks (DSBs) Induced by CYN, MCLR and Their Binary Mixture
2.3. The Influence of CYN, MCLR and Their Binary Mixture on the Cell-Cycle Progression
2.4. The Effects of CYN, MCLR and Their Mixtures on the mRNA Expression
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Chemicals
5.2. Cell Culture
5.3. Concentrations of CYN, MCLR and Their Combinations for Determining Cytotoxicity, Genotoxicity and mRNA Expression
5.4. Cytotoxicity Assay-MTS Test
5.5. Influence of Cyanotoxins on DNA Double-Strand Break Induction and Cell Cycle Determined by Flow Cytometry
5.6. Gene Expression Analysis by Quantitative Real-Time PCR (qPCR)
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Involved Pathway | Gene Symbol | Treatment (µg/mL) | 72 h Mean ± SD | Entrez Gene Name |
---|---|---|---|---|
Metabolism (activation/detoxification) | CYP1A1 | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 2.59 ± 0.89 1.03 ± 0.13 2.53 ± 1.53 48.87 ± 0.81 **** | Cytochrome P450 family 1 Subfamily A polypeptide 1 |
UGT1A1 | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 0.96 ± 0.45 0.97 ± 0.25 1.20 ± 0.69 2.94 ± 1.15 * | UDP glucuronosyltransferase 1 family polypeptide A1 | |
Immediate-early response/signaling | JUNB | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 1.22 ± 0.26 1.10 ± 0.24 1.16 ± 0.35 2.60 ± 0.45 *** | Jun B proto-oncogene |
DNA damage responsive | TP53 | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 0.76 ± 0.02 **** 0.96 ± 0.02 0.75 ± 0.06 **** 1.16 ± 0.04 ** | Tumor protein P53 |
MDM2 | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 1.25 ± 0.29 1.08 ± 0.08 1.11 ± 0.21 1.39 ± 0.08 | MDM2 oncogene, E3 ubiquitin-protein ligase | |
CDKN1A | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 2.44 ± 0.30 *** 1.12 ± 0.24 2.47 ± 0.43 *** 8.61 ± 0.16 **** | Cyclin-Dependent Kinase Inhibitor 1A | |
GADD45A | 0.5 CYN 1 MCLR 0.5 CYN + 1 MCLR B(a)P 30 µM | 3.16 ± 0.54 ** 1.11 ± 0.10 3.06 ± 0.89 ** 3.76 ± 0.75 *** | Growth arrest and DNA damage-inducible gene, alpha |
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Díez-Quijada, L.; Hercog, K.; Štampar, M.; Filipič, M.; Cameán, A.M.; Jos, Á.; Žegura, B. Genotoxic Effects of Cylindrospermopsin, Microcystin-LR and Their Binary Mixture in Human Hepatocellular Carcinoma (HepG2) Cell Line. Toxins 2020, 12, 778. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins12120778
Díez-Quijada L, Hercog K, Štampar M, Filipič M, Cameán AM, Jos Á, Žegura B. Genotoxic Effects of Cylindrospermopsin, Microcystin-LR and Their Binary Mixture in Human Hepatocellular Carcinoma (HepG2) Cell Line. Toxins. 2020; 12(12):778. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins12120778
Chicago/Turabian StyleDíez-Quijada, Leticia, Klara Hercog, Martina Štampar, Metka Filipič, Ana M. Cameán, Ángeles Jos, and Bojana Žegura. 2020. "Genotoxic Effects of Cylindrospermopsin, Microcystin-LR and Their Binary Mixture in Human Hepatocellular Carcinoma (HepG2) Cell Line" Toxins 12, no. 12: 778. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins12120778