Ratiometric fluorescence assay for γ-glutamyltranspeptidase detection based on a single fluorophore via analyte-induced variation of substitution

Hou, Xianfeng; Yu, Qingxiang; Zeng, Fang; Yu, Changmin; Wu, Shuizhu

doi:10.1039/c4cc00473f

Cited by 77 publications

(49 citation statements)

References 36 publications

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“…Here we have performed such an assay by using a mixed serum sample from healthy people and three individual serum samples from liver cancer patients. As shown in Table 1, the GGT level in the mixed healthy human serum is 66.50 U/L, which is in coincidence with the normal serum GGT level (5 -85 U/L), 15 whereas those in two serum samples from liver cancer patients are over 100 U/L, which exceeds the normal range. However, one of the serum samples from the liver cancer patients shows unexpectedly the same GGT level as that from the healthy people, namely obviously a false-negative result.…”

Section: ■ Results and Discussionmentioning

confidence: 83%

“…It was found that the fluorescence enhancement was directly proportional to the GGT concentration in the range of 1 -50 U/L, with a linear equation of ΔF = 16.1  C (U/L) + 7.8 (R = 0.996), where ΔF is the increase of fluorescence intensity with and without GGT (Figure 2). Moreover, a good linear fluorescence response of CV-Glu still holds even at lower concentrations (0 -100 mU/L) of GGT (Figure S11); in this case, the detection limit (3S/m, where S is the standard deviation of blank measurements, n = 11, and m is the slope of the linear equation) was determined to be 5.6 mU/L GGT, which is one of the most sensitive fluorescent probes 15. Then the Michaelis constant (K m ) of CV-Glu reacting with GGT was calculated by using Lineweaver-Burk plot (Figure S12in the Supporting Information).…”

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confidence: 99%

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Sensitive Fluorescence Probe with Long Analytical Wavelengths for γ-Glutamyl Transpeptidase Detection in Human Serum and Living Cells

Shi

Wang

et al. 2015

Anal. Chem.

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A new sensitive fluorescent probe with long analytical wavelengths for γ-glutamyl transpeptidase (GGT) assay has been developed by incorporating the γ-glutamyl group as a recognition unit into the fluorophore of cresyl violet (CV). The detection mechanism is based on the GGT-catalyzed cleavage of the γ-glutamyl group, followed by the release of CV, which leads to a distinct color change from light yellow to pink and a large fluorescence enhancement at 615 nm (λex = 585 nm). Under the optimized conditions, the fluorescence intensity of the probe is directly proportional to the activity of GGT in the range of 1-50 U/L with a detection limit of 5.6 mU/L. By virtue of its high sensitivity and long analytical wavelengths, the probe has been used to directly determine GGT in human serum samples from both healthy people and liver cancer patients, and the obtained results accord well with those acquired by commercial GGT fluorometric assay kit. The probe has also been employed to image endogenous GGT in living cells. Notably, with our probe the expression level of GGT in HepG2 cells under the action of sodium butyrate (an anticancer drug) was studied by fluorescence confocal microscopy, revealing that sodium butyrate can induce the upregulation of GGT in HepG2 cells in a dose- and time-dependent manner. This behavior of sodium butyrate has further been confirmed by lysate assay and inhibitor experiment. The proposed probe is rather simple and may find a wide use in the determination of GGT in clinical and biological samples.

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Section: ■ Results and Discussionmentioning

confidence: 83%

mentioning

confidence: 99%

Sensitive Fluorescence Probe with Long Analytical Wavelengths for γ-Glutamyl Transpeptidase Detection in Human Serum and Living Cells

Shi

Wang

et al. 2015

Anal. Chem.

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“…And this technique has been extensively applied in the research of various enzyme-catalyzed processes [51][52][53][54][55]. [56][57][58][59][60][61][62][63][64][65][66][67].…”

Section: Introductionmentioning

confidence: 99%

A fluorescent probe for alkaline phosphatase via excited state intramolecular proton transfer

Zeng

2015

Sensors and Actuators B: Chemical

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“…no. 23266; Thermo Fisher Scientific, Inc.) was used to detect caspase-3 and caspase-9 activity according to the manufacturer's protocol (19). In brief, vaginal epithelial cells were lysed in lysis buffer (Beyotime Biotechnology, Nantong, Jiangsu, China) and centrifuged at 18,000 x g for 10 min at 4˚C and supernatants were collected.…”

Section: Cell Viability Assaymentioning

confidence: 99%

Effects of siRNA-mediated suppression of HPV-11 L1 expression on the proliferation and apoptosis of vaginal epithelial cells

Zeng

Yang

Wang

et al. 2017

Experimental and Therapeutic Medicine

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Abstract. The aim of the present study was to investigate the effects of human papillomavirus (HPV) infection on the gynecological disease of vaginitis and to demonstrate how the small interfering RNA (siRNA) method may be used for HPV prevention in the clinic. Human vaginal epithelial cells were transfected with HPV-11 L1 expression vector and siRNA-HPV-11 L1 vectors and a control group was transfected with scrambled siRNA. Cell proliferation in each group was analyzed using the MTT assay and the expression of apoptosis-associated proteins was measured by western blot analysis. Compared with the control group, HPV-11 L1 mRNA and protein levels were significantly increased following transfection with the HPV-11 L1 expression vector in cells (P<0.05), but this result was significantly reversed by silencing of HPV-11 L1 (P<0.05). In addition, cell proliferation in the HPV-11 group was lower than that in the control group; however, cell proliferation was significantly increased in cells transfected with silenced L1 compared with that in the control group (P<0.05). Furthermore, silencing of HPV-11 L1 significantly decreased caspase-3 and caspase-9 expressions in cells, whereas the expression was increased in the HPV-11 L1 group (P<0.05). The present study suggested that siRNA-mediated silencing of HPV-11 L1 may have potential therapeutic applications for treating gynecological diseases associated with HPV-11 infection.

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Ratiometric fluorescence assay for γ-glutamyltranspeptidase detection based on a single fluorophore via analyte-induced variation of substitution

Cited by 77 publications

References 36 publications

Sensitive Fluorescence Probe with Long Analytical Wavelengths for γ-Glutamyl Transpeptidase Detection in Human Serum and Living Cells

Sensitive Fluorescence Probe with Long Analytical Wavelengths for γ-Glutamyl Transpeptidase Detection in Human Serum and Living Cells

A fluorescent probe for alkaline phosphatase via excited state intramolecular proton transfer

Effects of siRNA-mediated suppression of HPV-11 L1 expression on the proliferation and apoptosis of vaginal epithelial cells

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