Development of fluorescent dual-FRET probe for simultaneous detection of caspase-8 and caspase-9 activities and their relative quantification

Porubský, Martin; Řezníčková, Eva; Křupková, Soňa; Kryštof, Vladimı́r; Hlaváč, Jan

doi:10.1016/j.bioorg.2022.106151

Cited by 4 publications

(4 citation statements)

References 35 publications

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“…Apoptosis, a process of programmed cell death, occurs through both extrinsic and intrinsic pathways. While the extrinsic pathway is triggered by death receptors that result in caspase-8 activation, the intrinsic pathway is triggered by organelle injury leading to caspase-9 activation ( 32 , 33 ). Both pathways have been shown to activate the same downstream caspase-3 molecules, to eventually trigger cell apoptosis ( 25 , 34 , 35 ).…”

Section: Discussionmentioning

confidence: 99%

Solamargine induces apoptosis of human renal carcinoma cells via downregulating phosphorylated STAT3 expression

Huang,

Sun,

Ren

et al. 2023

Oncol Lett

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Solamargine (SM), an active compound derived from Solanum nigrum , triggers apoptosis and inhibits the metastatic and oxidative activities of various types of tumor cells. However, the effect of SM on human renal carcinoma cells remains unknown. In the present study, the molecular mechanisms underlying the antitumor effects of SM on ACHN and 786-O cells were elucidated. Specifically, MTT and colony formation assays were conducted to evaluate the impact of SM treatment on the proliferation of ACHN and 786-O cells, and flow cytometry was conducted to determine the influence of SM on the apoptosis rates of these cells. In addition, the expression of target proteins was determined by western blotting. The results revealed that SM not only inhibited cell viability but also promoted the apoptosis of ACHN and 786-O cells in a time- and dose-dependent manner. Moreover, treatment of ACHN and 786-O cells with SM significantly enhanced the caspase-3, caspase-8 and caspase-9 activities. Furthermore, SM downregulated the expression of phosphorylated signal transducer and activator of transcription-3 (p-STAT3) and Bcl-2 but increased the expression of cleaved caspase-3, −8, −9 and Bax. BAY2353, a p-STAT3 inhibitor, inhibited the viability of ACHN and 786-O cells, increased the expression of cleaved caspase-9 and Bax and decreased the expression of p-STAT3 and Bcl-2. Further experiments demonstrated that SM inhibited tumor growth in xenograft nude mice without causing specific toxicity to the major organs. Collectively, these findings indicated that SM not only inhibited the viability but also promoted the apoptosis of ACHN and 786-O cells, through a mechanism involving downregulation of p-STAT3 expression.

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Section: Discussionmentioning

confidence: 99%

Solamargine induces apoptosis of human renal carcinoma cells via downregulating phosphorylated STAT3 expression

Huang,

Sun,

Ren

et al. 2023

Oncol Lett

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“…After an emission spectrum for the time 0 min was measured, 10 μL of 1 mM HCl with or without protease in an appropriate concentration and 10 μL of water solution with or without hydrogen peroxide in a suitable amount were added, and the obtained sample in the covered fluorimeter cuvette was incubated at 37 °C for the time period of 45 min. The emission spectra upon uniform excitation with 425 nm were measured every five min at the times of 5,10,15,20,25,30,35,40, and 45 min.…”

Section: ■ Methods and Materialsmentioning

confidence: 99%

“…While quite a few studies describing multiplex protease sensing, − synchronous enzyme and pH monitoring, − and detection of hydrogen peroxide in combination with other nonenzymatic species − exist, to the best of our knowledge, there are only two known reports dealing with the simultaneous enzyme and hydrogen peroxide screening. Peng et al developed a dual-locked NIR fluorescence-based sensor for selective marking of melanoma cells, applying peroxide tyrosinase cascade activation of a fluorescence-inactive methylene blue borate derivative .…”

Section: Introductionmentioning

confidence: 99%

Novel Peptide-Based Fluorescent Probe for Simultaneous Sensing of Chymotrypsin and Hydrogen Peroxide

Milićević,

Hlaváč

2024

ACS Omega

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The developed multifunctional fluorescent probe enables the simultaneous detection of chymotrypsin as a model protease and hydrogen peroxide as a representative of reactive oxygen species (ROS) in biologically relevant concentration ranges. The chymotrypsin sensing is based on the cleavage of its selectively recognizable peptide sequence and the consequent disruption of FRET between coumarin (DEAC) and fluorescein (FL). Analogously, the presence of hydrogen peroxide causes the gradual degradation of the H 2 O 2 -labile benzopyrylium-coumarin (BC) dye. Considering the fluorescence emission responses of individual chymotrypsin-peroxide probe-attached fluorophores after their excitation at 425 nm, the sole presence of either chymotrypsin (50−1000 ng/ mL) or hydrogen peroxide (10−200 μM) in a sample could be unambiguously confirmed or refuted. In addition, reliable simultaneous detection and approximate quantification of both studied species in the concentration ranges of 100−1000 ng/ mL and 20−200 μM for chymotrypsin and H 2 O 2 , respectively, could be performed as well. The obtained results are summarized and visualized in the graphical models.

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“…The rhodamine derivative used as a uorophore for this probe has many merits of high quantum yield, great water-solubility, and a long emission. [39][40][41][42] Importantly, the positive charge carried by this uorophore endows it with the ability to target mitochondria. The additionally attached allyl moiety serves as the reaction site, and in response, the hydroxyl group is exposed, releasing a uorophore with a strong ICT structure, and the red strong uorescence is restored.…”

Section: Introductionmentioning

confidence: 99%

A long-wavelength mitochondria-targeted CO fluorescent probe for living cells and zebrafish imaging

Rong,

Liu,

et al. 2024

Anal. Methods

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Development of fluorescent dual-FRET probe for simultaneous detection of caspase-8 and caspase-9 activities and their relative quantification

Cited by 4 publications

References 35 publications

Solamargine induces apoptosis of human renal carcinoma cells via downregulating phosphorylated STAT3 expression

Solamargine induces apoptosis of human renal carcinoma cells via downregulating phosphorylated STAT3 expression

Novel Peptide-Based Fluorescent Probe for Simultaneous Sensing of Chymotrypsin and Hydrogen Peroxide

A long-wavelength mitochondria-targeted CO fluorescent probe for living cells and zebrafish imaging

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