ANALYTICAL CHEMISTRY
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2025
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IF 6.7
Lanlan Chen,Jingjing Lai,Siqi Dong,Wenjun Liu,Ximei Zhang,Huanghao Yang
Evaluating tumor radiosensitivity is beneficial for the prediction of treatment efficacy, customization of treatment plans, and minimization of side effects. Tracking the mitochondrial DNA (mtDNA) repair process helps to assess tumor radiosensitivity as mtDNA repair determines the fate of the cell under radiation-induced mtDNA damage. However, current probes developed to monitor levels of DNA repair enzymes suffered from complex synthesis, uncontrollable preparation, limited tumor selectivity, and poor organelle-targeting ability. Especially, the correlation between mtDNA repair activity and inherent radiosensitivity of tumors has not yet been explored. Here, we present a mitochondria-targeted DNA-based nanoprobe (TPP-Apt-tFNA) for in situ monitoring of the activity of the mtDNA repair enzyme and evaluating tumor radiosensitivity. TPP-Apt-tFNA consists of a DNA tetrahedral framework precisely modified with three functional modules on each of the three vertexes, that is, the tumor cell-targeting aptamer, the mitochondrion-targeting moiety, and the apurinic/apyrimidinic endonuclease 1 (APE1)-responsive molecule beacon. Once selectively internalized by tumor cells, the nanoprobe targeted the mitochondrion and specifically recognized APE1 to activate fluorescence, allowing the observation of mtDNA repair activity. The nanoprobe showed elevated APE1 levels in the mitochondria of tumor cells under oxidative stress. Moreover, the nanoprobe enabled the illumination of different levels of APE1-mediated mtDNA repair activity in different cell cycle phases. Furthermore, using the nanoprobe in vitro and in vivo, we found that tumor cells with high activity of mtDNA repair, which allowed them to recover from radiation-induced mtDNA lesions, had low sensitivity to radiation and an unsatisfactory radiotherapy outcome. Our work provides a new imaging tool for exploring the roles of mtDNA repair activity in diverse biological processes and for guiding tumor radiation treatment.
研究领域:
生物化学
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分子生物学
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医学影像学
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肿瘤学
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产品关联
DOI
PubMed
高影响因子
ACS Sensors
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2021
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IF 7.711
Chao Jiang,Fangfang Wang,Kexin Zhang,Tiantian Min,Desheng Chen,Yongqiang Wen
In the deoxyribonucleic acid (DNA) repair pathways, DNA repair enzymes have great significance for genomic integrity. As one important initiator of the base-excision repair pathway, the aberrant activity of uracil-DNA glycosylase (UDG) is closely associated with many diseases. Herein, we developed a simple distance-based device for visual detection of UDG activity using a load-free DNA hydrogel. The DNA hydrogel consists of polyacrylamide–DNA chains being bridged by a single-stranded DNA crosslinker containing a responsive uracil base site. UDG can recognize and remove the uracil, resulting in the cleavage effect of the DNA crosslinker strand with the assistance of endonuclease IV (Endo IV). Plugging one end of the capillary tube, the DNA hydrogel acting as a filter membrane separator would control molecules to flow into the tube. The integrity of the DNA hydrogel networks is affected by the excision of UDG. Therefore, taking full advantage of membrane filtration of the DNA hydrogel, the activity of UDG can be quantitatively detected via reading the distance of the red indicator solution in the capillary tube. Without any instruments and complicated procedures, this method realizes high sensitivity and specificity for the detection of UDG as low as 0.02 mU/mL and can even measure UDG in complex cell samples. Additionally, this method is simple, universal, and can be used to screen inhibitors, which shows great potential for point-of-care testing, clinical diagnosis, and drug discovery.
研究领域:
生物化学
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分子生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2019
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IF 6.393
Yu Ling,Jiao Zhou,Xiao Fang Zhang,Xiao Hu Wang,Nian Bing Li,Hong Qun Luo
As one of critical base excision repair enzymes, uracil-DNA glycosylase (UDG) can specifically repair uracil-induced DNA damage to maintain the genome integrity of organisms. Moreover, abnormal expression of UDG is related to various cancers and other serious diseases. Therefore, it is essential to accurately and sensitively monitor UDG activity. In this work, a ratiometric fluorescence method using poly-(thymine) DNA template copper nanoclusters (Cu NCs) and 4′,6-diamidino-2-phenylindole (DAPI) as the output signals is developed for simple, selective, and sensitive detection of UDG. A double-stranded DNA (dsDNA) consisting of a uracil-containing single chain and a trigger sequence is designed as a substrate dsDNA. Meanwhile, another dsDNA comprising 30 pairs of A-T bases ((AT) 30 dsDNA) is designed to be partially complementary to the trigger chain. The (AT) 30 dsDNA itself has no blunt or recessed 3′ end and cannot be cut by exonuclease III (Exo III). With the Exo III-assisted amplification, adding UDG results in the success synthesis of red-emissive Cu NCs, while the dsDNAs in the system are hydrolyzed and the fluorescence signal of DAPI is weak. In contrast, without addition of UDG, the fluorescence intensity of Cu NCs is low, while the DAPI emits significantly enhanced fluorescence signal. The proposed method presents a detection limit of 5.0 × 10 −5 U mL −1 and can be applied to the HeLa cell lysate sample with satisfactory results. Additionally, the kinetic parameter is studied and the UDG inhibition effect is evaluated. In summary, this work provides a potential platform for DNA repair enzyme-related analysis and clinical diagnosis.
研究领域:
生物化学
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分子生物学
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癌症研究
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分析化学
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产品关联
DOI
高影响因子
ELECTROCHIMICA ACTA
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2019
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IF 5.383
Xia-Xia Liang,Lei Qian,Rong-Fu Huang
Endogenous cell metabolism and chemical reactions can induce oxidative DNA damage, which is involved in the development of different diseases. In this work, a label-free electrochemiluminescence (ECL) assay was proposed for the sensitive detection of oxidative DNA damage in an assembled DNA film. We employed the formamidopyrimidine-DNA glycosylase (Fpg) to convert DNA oxidative nucleobases to strand break, allowing to detect the DNA damage inducing minimal change to the helical structure of DNA. A “light-switch” DNA intercalator, [Ru(bpy) 2 (dppz)] 2+ (bpy = 2, 2 ′ -bipyridine, dppz = dipyrido[3, 2-a:2 ′ , 3 ′ -c]phenazine) was introduced as an ECL signal reporter to detect the DNA strand breaks. The damaged DNA film on the reduced graphene oxide and gold nanoparticles (r-GO@Au) modified ITO electrode bound less ECL indicator than the intact film and accompanied by a drop in ECL intensity, which were also confirmed by gel electrophoresis and fluorescence measurements. As a result, a Fe 2+ -mediated Fenton reaction with as low as 10 nM Fe 2+ and 40 nM H 2 O 2 induced DNA damage can be detected, 100-fold lower than the concentration measured without Fpg. To demonstrate its universality, the successful detection of DNA damage induced by Rose Bengal under irradiation was also achieved. The proposed strategy shows its potential practicality for the sensitive and rapid assessment of the genotoxicity of environmental pollutants .
研究领域:
生物化学
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分析化学
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分子生物学
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环境科学
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产品关联
DOI
高影响因子
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