A cathodic photoelectrochemical immunoassay with dual signal amplification for the ultrasensitive detection of DNA damage biomarkers

Zhang, Bihong; Li, Fangfang; Shen, Linyu; Chen, Lu; Xia, Zhiqiang; Ding, Jin-Jian; Li, Minjie; Guo, Liang-Hong

doi:10.1016/j.bios.2022.115052

Cited by 4 publications

(2 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…γ H2A.X plays a crucial role in early DNA damage response and is widely regarded as the primary indicator for measuring DSB levels. 64 , 65 RNF8 accumulates rapidly at the damaged sites during the repair and facilitates recruitment of repair proteins such as p53-binding protein 1 (53BP1) and breast cancer susceptibility protein 1 (BRCA1) to the damaged chromatin by regulating ubiquitination of histone H2A and H2AX, thus being indispensable for the repair process. 61 , 62 , 66 In our prior study, we employed RNF8 knockout model mice and observed that RNF8 is essential for DNA damage repair in mouse neurons.…”

Section: Discussionmentioning

confidence: 99%

SiRNF8 Delivered by DNA Framework Nucleic Acid Effectively Sensitizes Chemotherapy in Colon Cancer

Guo,

Song,

Tian

et al. 2024

IJN

View full text Add to dashboard Cite

Background The evident side effects and decreased drug sensitivity significantly restrict the use of chemotherapy. However, nanoparticles based on biomaterials are anticipated to address this challenge. Methods Through bioinformatics analysis and colon cancer samples, we initially investigated the expression level of RNF8 in colon cancer. Next, we constructed nanocarrier for delivering siRNF8 based on DNA tetrahedron (si-Tet), and Doxorubicin (DOX) was further intercalated into the DNA structure (si-DOX-Tet) for combination therapy. Further, the effects and mechanism of RNF8 inhibition on the sensitivity of colon cancer cells to DOX chemotherapy have also been studied. Results RNF8 expression was increased in colon cancer. Agarose gel electrophoresis, transmission electron microscopy, and size distribution and potential analysis confirmed the successful preparation of the two nanoparticles, with particle sizes of 10.29 and 37.29 nm, respectively. Fluorescence imaging reveals that the carriers can be internalized into colon cancer cells and escape from lysosomes after 12 hours of treatment, effectively delivering siRNF8 and DOX. Importantly, Western blot analysis verified treatment with 50nM si-Tet silenced RNF8 expression by approximately 50% in colon cancer cells, and combined treatment significantly inhibited cell proliferation. Furthermore, the CCK-8 assay demonstrated that si-Tet treatment enhanced the sensitivity of colon cancer cells to the three chemotherapeutic drugs. Significant more DNA damage was detected after treatment with both si-Tet or si-DOX-Tet. Further flow cytometry analysis revealed that si-DOX-Tet treatment led to significantly more apoptosis, approximately 1.6-fold higher than treatment with DOX alone. Mechanistically, inhibiting RNF8 led to decreased ABCG2 expression and DOX efflux, but increased DNA damage, thereby enhancing the chemotherapeutic effect of DOX. Conclusion We have successfully constructed si-DOX-Tet. By inhibiting the expression of RNF8, it enhances the chemotherapy sensitivity of DOX. Therefore, this tetrahedral FNA nanocarrier offers a new approach for the combined treatment of colon cancer.

show abstract

Section: Discussionmentioning

confidence: 99%

SiRNF8 Delivered by DNA Framework Nucleic Acid Effectively Sensitizes Chemotherapy in Colon Cancer

Guo,

Song,

Tian

et al. 2024

IJN

View full text Add to dashboard Cite

show abstract

“…Antibodies or commercial kits for γH2AX detection are available and suitable for flow cytometry as well as for microscopic analysis and other novel immuno-based methods [11]. Flow cytometry only allows us to measure overall fluorescence, whereas γH2AX detection by immunofluorescence microscopy is based either on the foci detection per cell (using a confocal microscope) or on the quantification of the overall intensity from images of lower magnification [10], the second mentioned approach being more suitable for highthroughput analysis in a 96-well plate format.…”

Section: Introductionmentioning

confidence: 99%

In Vitro High-Throughput Genotoxicity Testing Using γH2AX Biomarker, Microscopy and Reproducible Automatic Image Analysis in ImageJ—A Pilot Study with Valinomycin

Křížkovská

Schätz

Lipov

et al. 2023

Toxins

View full text Add to dashboard Cite

(1) Background: The detection of DNA double-strand breaks in vitro using the phosphorylated histone biomarker (γH2AX) is an increasingly popular method of measuring in vitro genotoxicity, as it is sensitive, specific and suitable for high-throughput analysis. The γH2AX response is either detected by flow cytometry or microscopy, the latter being more accessible. However, authors sparsely publish details, data, and workflows from overall fluorescence intensity quantification, which hinders the reproducibility. (2) Methods: We used valinomycin as a model genotoxin, two cell lines (HeLa and CHO-K1) and a commercial kit for γH2AX immunofluorescence detection. Bioimage analysis was performed using the open-source software ImageJ. Mean fluorescent values were measured using segmented nuclei from the DAPI channel and the results were expressed as the area-scaled relative fold change in γH2AX fluorescence over the control. Cytotoxicity is expressed as the relative area of the nuclei. We present the workflows, data, and scripts on GitHub. (3) Results: The outputs obtained by an introduced method are in accordance with expected results, i.e., valinomycin was genotoxic and cytotoxic to both cell lines used after 24 h of incubation. (4) Conclusions: The overall fluorescence intensity of γH2AX obtained from bioimage analysis appears to be a promising alternative to flow cytometry. Workflow, data, and script sharing are crucial for further improvement of the bioimage analysis methods.

show abstract

Near-infrared light-driven lab-on-paper cathodic photoelectrochemical aptasensing for di(2-ethylhexyl)phthalate based on AgInS2/Cu2O/FeOOH photocathode

Yang,

Tu,

Hao

et al. 2024

Talanta

View full text Add to dashboard Cite

A cathodic photoelectrochemical immunoassay with dual signal amplification for the ultrasensitive detection of DNA damage biomarkers

Cited by 4 publications

References 21 publications

SiRNF8 Delivered by DNA Framework Nucleic Acid Effectively Sensitizes Chemotherapy in Colon Cancer

SiRNF8 Delivered by DNA Framework Nucleic Acid Effectively Sensitizes Chemotherapy in Colon Cancer

In Vitro High-Throughput Genotoxicity Testing Using γH2AX Biomarker, Microscopy and Reproducible Automatic Image Analysis in ImageJ—A Pilot Study with Valinomycin

Near-infrared light-driven lab-on-paper cathodic photoelectrochemical aptasensing for di(2-ethylhexyl)phthalate based on AgInS2/Cu2O/FeOOH photocathode

Contact Info

Product

Resources

About