International Journal of Peptide Research and Therapeutics
·
2021
·Vol. 27
·期 2
·1343-1350
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IF 1.931
Zhao Yi,Sun Guangming,Yuan Libo,Li Jinlei,Lu Kui
Breast cancer susceptibility gene 1 (BRCA1) is an important tumor suppressor gene in the human which associated with a variety of tumor suppressor genes to inhibit the growth of malignant tumors. Breast cancer is closely related to BRCA1 mutations in humans. Four BRCA1 (856-871) mutants were screened that can interact well with RAD51 (158-180) using computer to simulate mutations at Y 856, R 866 and Q 867. These mutants were synthesized by using Fmoc solid-phase synthesis, and purified by liquid chromatography. According to the results of fluorescence spectroscopy, R866W has the strongest binding ability for the above peptides and RAD51 (158-180). Based on the circular dichroism spectrum results, the sequence of the interaction ability of the above peptides on the secondary structure of RAD51 (158-180) is Y856P, Q867C > Y856P, R866W ≈ R866W > Y856P > BRCA1. All the spectral results show that R866W have the strongest interaction ability with RAD51(158-180).
研究领域:
分子生物学
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肿瘤学
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生物化学
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高影响因子
International Journal of Peptide Research and Therapeutics
·
2019
·Vol. 26
·期 1
·121-128
·
IF 1.219
Wang Mengwei,Lv Mingxiu,Lu Kui,Liu Guangbin,Mai Wenpeng,Yu Bo,Lou Yudan
Six breast cancer susceptibility gene 1 (BRCA1) peptide fragments were designed by removing N-terminal amino acid residues of BRCA1(846–871). Peptide fragments BRCA1(846–871), BRCA1(852–871), BRCA1(854–871), BRCA1(856–871), BRCA1(857–871) and BRCA1(861–871) were obtained by Fmoc solid-phase synthesis, purified by reversed-phase high-performance liquid chromatography and characterized by electrospray ionization mass spectrometry. The interaction between BRCA1 fragment peptides and RAD51(181–200) target peptides were investigated using circular dichroism (CD) spectroscopy, fluorescence spectroscopy, and microscale thermophoresis (MST). The results of CD spectroscopy showed that the secondary structures of RAD51(181–200) were altered after the addition of BRCA1 peptide fragments. The fluorescence results proved that all six BRCA1 peptide fragments BRCA1(846–871), BRCA1(852–871), BRCA1(854–871), BRCA1(856–871), BRCA1(857–871) and BRCA1(861–871) could bind to RAD51(181–200) independently, their quenching processes were static, and the binding constant of peptide fragments with RAD51(181–200) were 4.01 × 10 5 L mol −1 , 7.22 × 10 5 L mol −1 , 7.84 × 10 5 L mol −1 , 1.1 × 10 4 L mol −1 , 6.19 × 10 3 L mol −1 , 6.18 × 10 3 L mol −1 . The results of MST displayed that BRCA1(854–871) could combine with RAD51(181–200), and the dissociation constant K d was 126.44 µM. Compared with other five peptide fragments, the interaction between BRCA1(854–871) and RAD51(181–200) was much stronger.
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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高影响因子
JOURNAL OF THE BRAZILIAN CHEMICAL SOCIETY
·
2023
·Vol. 34
·期 6
·857-865
·
IF 1.4
Hu Jing,Yang Tong,Li Zhengjia,Xiao Bo,Yu Peng
The mutations of breast cancer susceptibility gene 1 (BRCA1) play an important role in inherited breast cancers. Thus, the sensitive assay of the BRCA1 gene is extremely important for disease diagnosis and human health. Herein, a fast, sensitive and selective assay technique has been constructed based on fluorescent copper nanoclusters (CuNCs). The CuNCs were successfully formed with poly(AT-TA) double stranded DNA (dsDNA) as template. In the absence of the target, a strong red emission was observed under 365 nm ultraviolet (UV) lamp and a big fluorescence response was obtained. However, in the presence of BRCA1 gene, there was only weak red emission and a low response signal was produced because of the target-proximity induced quenching of the fluorescence of CuNCs. The linear range for the BRCA1 gene assay was 2-600 nM, and the limit of detection was 2 nM. The assay technique showed good selectivity, good stability and satisfactory recoveries for the detection of BRCA1 gene in diluted serum samples. Moreover, by integrating a UV lamp and a smartphone, the fluorescence sensor would be transferred to a microfluidic chip, providing a prospective application in point-of-care for monitoring breast cancer risk.
研究领域:
分子生物学
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医学诊断
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纳米技术
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Bo Dong,Kaixuan Nie,Huanhuan Shi,Lemeng Chao,Mingyang Ma,Fengxiao Gao,Bo Liang,Wei Chen,Mengqiu Long,Zhengchun Liu
Peptide nucleic acids array technology is a method of greatly increasing the throughput of laboratory processes to efficiently perform large-scale genetic tests. Diethylene glycol-containing chiral γPNA (miniPEG-γPNA) is considered to be the best PNA derivative and one of the best candidates for gene detection, because it can hybridize DNA with greater affinity and sequence selectivity than DNA and ordinary aminoethylglycyl PNA ( aeg PNA). Herein, miniPEG-γPNA probes are synthesized by 9-fluorenylmethyloxycarbonyl (Fmoc) solid phase peptide synthesis (SPPS) in a mild condition, and a new biochip fabrication method “Film-Spotting” is invented, by which γPNA arrays with regular pattern, uniform luminance, and very low fluorescence background are obtained easily and cheaply. The miniPEG-γPNA array can effectively distinguish the full matched and mismatched targets in SSarc buffer, serum and urine, and the detection limit of complementary DNA is less than 5.97 nM. A miniPEG-γPNA array for BRCA1 gene mutation (3099delT) detection is also fabricated with a very good detection performance. This work provides an effective avenue for the diagnosis of breast cancer biomarker and expands the application of miniPEG-γPNA in the field of biochip.
研究领域:
分子生物学
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遗传学
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生物技术
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癌症诊断
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Chengchen Tan,Guoyang Xie,Song Wu,Cailu Song,Jinhui Zhang,Xinyao Yi,Jianxiu Wang,Hailin Tang
Breast cancer is reported to be one of the most lethal cancers in women, and its multi-target detection can help improve the accuracy of diagnosis. In this work, a cluster regularly interspaced short palindromic repeats (CRISPR)-Cas13a/Cas12a-based system was established for the simultaneous fluorescence detection of breast cancer biomarkers circROBO1 and BRCA1. CRISPR-Cas13a and CRISPR-Cas12a were directly activated by their respective targets, resulting in the cleavage of short RNA and DNA reporters, respectively, thus the signals of 6-carboxyfluorescein (FAM) and 6-carboxy-xrhodamine (ROX) were restored. As the fluorescence intensities of FAM and ROX were dependent on the concentrations of circROBO1 and BRCA1, respectively, synchronous fluorescence scanning could achieve one-step detection of circROBO1 and BRCA1 with detection limits of 0.013 pM and 0.26 pM, respectively. The system was highly sensitive and specific, holding high diagnostic potential for the detection of clinical samples. Furthermore, the competing endogenous RNA mechanism between circROBO1 and BRCA1 was also explored, providing a reliable basis for the intrinsic regulatory mechanism of breast cancer.
研究领域:
分子生物学
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癌症研究
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生物技术
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医学诊断
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产品关联
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2020
·
IF 4.722
Jiasheng Wang,Dongwei Wang,Ni Hui
The application of polypeptides in bio-interfaces and biosensors is of great interest because polypeptides are biocompatible and easy to design. A novel polymer nanocomposite was prepared by the electropolymerization of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) with a newly designed polypeptide. The nanocomposite polypeptide doped PEDOT (PEDOT/PEP), with a 3D microporous network structure, large surface area and excellent antifouling ability, was utilized for the attachment of BRCA1 complementary oligonucleotides to construct a DNA biosensor. The fabricated DNA biosensor showed favorable selectivity (with a detection limit of 0.0034 pM) and high sensitivity. The biosensor was also capable of detecting the target DNA (BRCA1) in 1% (V/V) human serum samples. The combination of a conducting polymer PEDOT with an antifouling and biocompatible polypeptide demonstrates a new method for preparing electrochemical sensors, that are capable of detecting targets in complex biological samples without strong nonspecific protein adsorption.
研究领域:
生物传感器
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材料科学
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癌症研究
·
电化学
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DOI
PubMed
高影响因子
ANALYST
·
2024
·
IF 3.6
Chengchen Tan,Xiaolong Yan,Xingchang Lu,Jianxiu Wang,Xinyao Yi
Breast cancer, as a malignant tumor with the highest incidence in the world, poses a great threat to human life and health. Early diagnosis of breast cancer may help enhancing the survival rate. In this work, a colorimetric and fluorescent dual-mode biosensor based on CRISPR-Cas12a system was constructed for the detection of breast cancer biomarker BRCA1. The intact G4 DNA could form DNAzyme in the assistance of K+ and hemin, catalyzed the oxidation of o-phenylenediamine (OPD) under the assistance of hydrogen peroxide (H2O2), and generate the oxidation product 2,3-diaminophenazine (DAP) with obvious absorption and fluorescence peaks. The presence of the target BRCA1 can activate the trans-cleavage activity of CRISPR-Cas12a, cleaving the G4 DNA and inhibiting the catalytic oxidation of OPD. The target BRCA1 was quantitatively determined by detecting both the absorbance and fluorescence intensity of DAP. Compared with single signal analysis, dual-signal output mode provided more reliable detection results. The detection limits were calculated to be 0.615 nM for colorimetric method and 0.289 nM for fluorescence method. The dual-mode biosensor showed good selectivity for BRCA1 and can resist the interference of complex substrates, which has great potential in biomedical detection.
研究领域:
生物医学工程
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分子生物学
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癌症研究
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生物技术
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Boyuan Pan,Mingxiu Lv,Heng Du,Dongxin Zhao,Kui Lu
Various peptide drugs have entered the market with the development of molecular biology. Peptide drugs are used for treat diseases such as diabetes, breast cancer, and HIV infection. In this study, three nicotinamide–modified peptides were synthesized by modifying the N–terminus of BRCA1 (856–871, Y856R, K862Y, R866W) peptide with three nicotinic acid derivatives using solid–phase peptide synthesis. The results of calf thymus DNA (ctDNA) binding activity indicated that binding constants of BRCA1 (856–871, Y856R, K862Y, R866W) (P0) and three nicotinamide–modified peptides (P1, P2, and P3) to ctDNA were 1.89 × 10 3 , 2.97 × 10 4 , 7.61 × 10 4 , and 8.09 × 10 4 L·mol −1 , respectively. The binding affinity of the modified peptides was superior to that of BRCA1 (856–871, Y856R, K862Y, R866W). ΔH θ < 0 and ΔS θ < 0 indicated that van der Waals force and hydrogen bond contributed most to peptide–ctDNA binding. Results obtained by Circular dichroism (CD) indicated that peptide binding interaction led to conformational changes in ctDNA. Ultraviolet–visible (UV) spectroscopy, ethidium bromide (EB) competition experiments, DNA melting experiments, and viscosity measurements verified that peptides interacted with ctDNA via groove binding. Ionic strength experiments manifested that electrostatic binding was also involved in peptide–ctDNA binding.
研究领域:
分子生物学
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药物化学
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生物化学
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生物物理学
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DOI
PubMed
高影响因子
International Journal of Peptide Research and Therapeutics
·
2019
·Vol. 26
·期 4
·1991-1999
·
IF 1.219
Cao Mengjie,Lu Kui,Liu Guangbin,Zhao Dongxin,Qi Jianzhao
Affecting the repair function of RAD51 can effectively reverse tumor resistance to chemotherapy and radiotherapy. There is interaction between RAD51 181−200 and BRCA1 846−871 . Using the computer to simulate single mutation of lysine at 862 position of BRCA1 846−871 , six BRCA1 846−871 mutants with significant interaction with RAD51 181−200 were screened out. These mutant peptides were obtained by 9-fluorenylmethyloxycarbonyl (Fmoc) solid-phase peptide synthesis (SPPS) method. All peptides were purified and characterized by liquid chromatography and ESI-MS. In this study, the interaction between RAD51 181−200 and BRCA1 846−871 or mutant peptides was investigated using circular dichroism spectroscopy (CD) and fluorescence spectroscopy. The CD results shown that the secondary structure change of RAD51 181−200 induced by BRCA1 846−871 , BRCA1 846−871 K862F ( F ), BRCA1 846−871 K862G ( G ), BRCA1 846−871 K862E ( E ) and BRCA1 846−871 K862N ( N ) was obvious, which the α-helix content of RAD51 181−200 decreased from 58.3–36.1%, 58.3–36.2%, 58.3–36.4%, 58.3–36.5% and 58.3–36.0%, respectively. While, the addition of BRCA1 846−871 K862P ( P ) and BRCA1 846−871 K862Y ( Y ) hardly changed the α-helix content of RAD51 181−200 . From the fluorescence results, the binding constants of F and Y are 1.00 × 10 5 M −1 and 1.16 × 10 5 M −1 respectively, which were both larger than that of BRCA1 846−871 (9.01 × 10 2 M −1 ). The binding constant of RAD51 181−200 with E (7.45 × 10 3 M −1 ) is slightly higher than that of BRCA1 846−871 . The binding constants of RAD51 181−200 with P (1.24 × 10 3 M −1 ) and N (1.26 × 10 3 M −1 ) are similar to that of BRCA1 846−871 . However, the K b value of G (1.51 × 10 2 M −1 ) is less than that of BRCA1 846−871 . Comparing all spectral data, F is the most significant one interacting with RAD51 181−200 . These findings could provide direction for peptide processing and application.
研究领域:
生物化学
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分子生物学
·
肿瘤学
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DOI
高影响因子
Translational Andrology and Urology
·
2022
·
IF 2.479
Shujun Sun,Kehua Jiang,Jin Zeng
Background To investigate the potential mechanisms of chemoradiotherapy resistance in patients with bladder cancer. Methods We assessed three bladder cancer cell lines (5637, J82, and TCCSUP) for their sensitivity to chemoradiotherapy. Reverse transcription quantitative PCR (RT-qPCR) was used to detect the expression of specific genes after chemoradiotherapy or combined with olaparib. The Genome Cancer Atlas (TGCA) database was used to analyze possible radioresistance-related genes and the relationship between their expression and bladder cancer survival and prognostic indicators. Results The 5637 cell line showed the most significant sensitivity to chemoradiotherapy. The expression levels of DNA damage repair genes in 5637 cells did not significantly change after chemoradiotherapy. In contrast, the expression levels of BRCA1 and RBBP8 genes significantly increased in J82 and TCCSUP cells after chemoradiotherapy. After combination with olaparib, which is a poly (ADP-ribose) polymerase inhibitor that initiates DNA repair, 5637 cells were significantly inhibited by chemoradiotherapy. However, chemoradiotherapy inhibition on J82 cells was weakened when combined with olaparib. A high reactive expression of BRCA1 and RBBP8 after combination with olaparib suggested that olaparib was ineffective because it did not induce synthetic lethality in which inhibiting PARP by olaparib coincided with suppression of BRCA1/2 expression result in cancer cell death. Conclusions The expression levels of RBBP8 and BRCA1 genes were associated with sensitivity to radiotherapy and chemotherapy in bladder cancer, and an increase in reactive expression after treatment led to worse sensitivity. Therefore, the reactive expression levels of BRCA1 and RBBP8 after chemoradiotherapy may be useful in evaluating the efficacy of olaparib combination therapy.
研究领域:
肿瘤学
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分子生物学
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医学遗传学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Wenting Wang,Xiaojian Fan,Shenghao Xu,Jason J. Davis,Xiliang Luo
A label-free and low fouling biosensor based on functional polyethylene glycols selective for breast cancer susceptibility gene (BRCA1) is reported. Sensory interfaces were prepared through the modification of a glassy carbon electrode with highly cross-linked polyethylene glycol (PEG) film containing amine groups, followed by the self-assembly of gold nanoparticles and the immobilization of BRCA1 complementary single-strand 19-mer oligonucleotides . In the presence of a specific BRCA1 sequence capture and hybridization results in interfacial change sensitively monitored using electrochemical impedance spectroscopy. The combined utilization of a PEG polymer film and gold nanoparticle mixed interface enables very high levels of sensitivity and a highly effective assaying in patient samples. Assay linear range was from 50.0 fM to 1.0 nM, with a limit of detection of 1.72 fM. Furthermore, this label-free DNA sensor has been used for assaying BRCA1 in serum samples, showing its feasible potential for diagnostic applications in clinical analysis of breast cancer gene BRCA1. Foreseeable, this sensor made on this basis undoubtedly provide the most effective and sensitive detection for BRCA1.
研究领域:
生物传感器
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癌症诊断
·
纳米技术
·
电化学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Guixiang Wang,Rui Han,Xiaoli Su,Yinan Li,Guiyun Xu,Xiliang Luo
Zwitterionic peptides were anchored to a conducting polymer of citrate doped poly(3,4-ethylenedioxythiophene) (PEDOT) via the nickel cation coordination, and the obtained peptide modified PEDOT, with excellent antifouling ability and good conductivity , was further used for the immobilization of a DNA probe to construct an electrochemical biosensor for the breast cancer marker BRCA1 . The DNA biosensor was highly sensitive (with detection limit of 0.03 fM) and selective, and it was able to detect BRCA1 in 5% (v/v) human plasma with satisfying accuracy and low fouling. The marriage of antifouling and biocompatible peptides with conducting polymers opened a new avenue to construct electrochemical biosensors capable of assaying targets in complex biological media with high sensitivity and without biofouling.
研究领域:
生物传感器
·
电化学
·
材料科学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）