Development of a Repair Enzyme Fluorescent Probe to Reveal the Intracellular DNA Damage Induced by Benzo[a]pyrene in Living Cells

Abstract

Pollutant exposure causes a series of DNA damage in cells, resulting in the initiation and progression of diseases and even cancers. An investigation of the DNA damage induced by pollutants in living cells is significant to evaluate the cytotoxicity, genotoxicity, and carcinogenicity of environmental exposure, providing critical insight in the exploration of the etiologies of diseases. In this study, we develop a repair enzyme fluorescent probe to reveal the DNA damage caused by an environmental pollutant in living cells by single-cell fluorescent imaging of the most common base damage repair enzyme named human apurinic/apyrimidinic endonuclease 1 (APE1). The repair enzyme fluorescent probe is fabricated by conjugation of an APE1 high affinity DNA substrate on a ZnO2 nanoparticle surface to form a ZnO2@DNA nanoprobe. The ZnO2 nanoparticle serves as both a probe carrier and a cofactor supplier, releasing Zn2+ to activate APE1 generated by pollutant exposure. The AP-site in the DNA substrate of the fluorescent probe is cleaved by the activated APE1, releasing fluorophore and generating fluorescent signals to indicate the position and degree of APE1-related DNA base damage in living cells. Subsequently, the developed ZnO2@DNA fluorescent probe is applied to investigate the APE1-related DNA base damage induced by benzo[a]pyrene (BaP) in living human hepatocytes. Significant DNA base damage by BaP exposure is revealed, with a positive correlation of the damage degree with exposure time in 2–24 h and the concentration in 5–150 μM, respectively. The experimental results demonstrate that BaP has a significant effect on the AP-site damage, and the degree of DNA base damage is time-dependent and concentration-dependent.

Strategic Assessment of Boron-Enriched Carbon Dots/Naproxen: Diagnostic, Toxicity, and In Vivo Therapeutic Evaluation

“…The study employed a mouse model induced with benzo­( a )­pyrene compounds (BAP) to assess further the therapeutic effectiveness of developing cancer. BAP, a polycyclic aromatic hydrocarbon (PAH), is known for its exceptionally high carcinogenic potential. , This propensity arises from the metabolic conversion of PAH compounds in the body into diol epoxides, , which subsequently bind to DNA and initiate the carcinogenic process. The results demonstrated that the B-CDs/Naproxen group exhibited a significant decrease in malignancy diameter throughout the study, while the control group showed a notable increase in cancer size volume.…”

Nanoconfined Silver Nanoclusters Combined with X-Shaped DNA Recognizer-Triggered Cascade Amplification for Electrochemiluminescence Detection of APE1

“…However, the recognition efficiency was constrained by the limited local concentration of the AP site on DNA hairpin recognizers. Researchers found that improving the local concentration of hairpins was a very effective approach for boosting reaction kinetics. , For example, Deng’s group developed the spherical nucleic acids by attaching AP site modified DNA hairpins on the surface of the ZnO 2 nanoparticles, allowing for highly sensitive and selective detection of APE1 . However, the DNA hairpins assembled on the ZnO 2 surface were disordered and time-consuming, making it difficult to avoid invalid collisions and steric hindrance .…”