Evaluation of the cytotoxic effect of Ly2109761 on HeLa cells using the xCELLigence RTCA system

Çeti̇n, İdi̇l; Topçul, Mehmet

doi:10.3892/ol.2018.9556

Cited by 10 publications

(7 citation statements)

References 18 publications

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“…In the present study, we confirmed by RTCA and MTT that different concentrations of isoimperatorin had significant inhibitory effects on nasopharyngeal carcinoma CNE2 cell proliferation and that the inhibitory effect increased as the concentration increased. Prior studies as well as the RTCA Evidence-Based Complementary and Alternative Medicine results presented here suggest that isoimperatorin may cause DNA damage resulting in loss of the cell's ability to undergo replication and transcription [22,23]. Flow cytometry analysis showed that, compared with the solvent group, isoimperatorin induced the CNE2 cell apoptosis.…”

Section: Discussionmentioning

confidence: 53%

Isoimperatorin Induces Apoptosis of Nasopharyngeal Carcinoma Cells via the MAPK/ERK1/2 Signaling Pathway

Liu

et al. 2020

Evidence-Based Complementary and Alternative Medicine

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Objective. To investigate the effect of isoimperatorin on nasopharyngeal carcinoma CNE2 cell apoptosis and the role of the MAPK/ERK1/2 signaling pathway in inducing apoptosis. Methods. Real-time cellular analysis technology (RTCA) and MTT were used to detect cell proliferation; Annexin V-FITC/PI dual-fluorescence flow cytometry analysis, Hoechst 33342 staining, and mitochondrial membrane potential detection kit were used to detect cell apoptosis; western blot was used to detect protein expression. Results. Different concentrations of isoimperatorin (10 μM, 20 μM, 30 μM, and 40 μM) significantly inhibited the nasopharyngeal carcinoma CNE2 cell proliferation, and 48 h later, the inhibitory effect of the 40 μM treatment was significantly higher than that of 10 μM and 20 μM. Treatment for 48 h significantly induced nasopharyngeal carcinoma cell apoptosis and resulted in nuclear pyknosis and fragmentation. At the same timepoint, the expression levels of proliferation-related protein PCNA as well as antiapoptosis proteins XIAP, survivin, and Bcl-2 were decreased by drug treatment, the expression level of proapoptosis protein Bax was increased, and the expression of the key MAPK/ERK1/2 signaling pathway proteins p-c-Raf, p-MEK, and p-ERK1/2 of were decreased. After activation of the MAPK/ERK1/2 signaling pathway by isoprenaline hydrochloride (ISO), the efficacy of isoimperatorin to downregulate p-c-Raf, p-MEK, and p-ERK1/2 expressions in the MAPK/ERK1/2 signaling pathway and proliferation-related protein PCNA as well as antiapoptosis proteins XIAP, surviving, and Bcl-2 was reduced compared with that of isoimperatorin alone, the effect of upregulating the proapoptotic protein Bax was reduced, and the apoptosis rate was also decreased. Conclusion. Isoimperatorin can induce nasopharyngeal carcinoma CNE2 cell apoptosis through the MAPK/ERK1/2 signaling pathway.

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Section: Discussionmentioning

confidence: 53%

Isoimperatorin Induces Apoptosis of Nasopharyngeal Carcinoma Cells via the MAPK/ERK1/2 Signaling Pathway

Liu

et al. 2020

Evidence-Based Complementary and Alternative Medicine

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“…RTCA can monitor infected HeLa cells inoculated onto a gold matrix by measuring the cell index (CI), an indicator that reflects cellular changes, including cell adhesion and cell number ( Roshan Moniri et al., 2015 ; Cetin and Topcul, 2019 ). The CI readouts of the WT strain infected HeLa cells were significantly lower than those of the mutant strains infected cells infected with the mutant strains from 2h to 9h post infection (12h to 21h in the RTCA assay) ( Figure 2 ).…”

Section: Resultsmentioning

confidence: 99%

Attenuation of Yersinia pestis fyuA Mutants Caused by Iron Uptake Inhibition and Decreased Survivability in Macrophages

Chen

Song

Chen

et al. 2022

Front. Cell. Infect. Microbiol.

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Yersinia pestis is the etiological agent of plague, a deadly infectious disease that has caused millions of deaths throughout history. Obtaining iron from the host is very important for bacterial pathogenicity. Y. pestis possesses many iron uptake systems. Yersiniabactin (Ybt) plays a major role in iron uptake in vivo and in vitro, and in virulence toward mice as well. FyuA, a β-barrel TonB-dependent outer membrane protein, serves as the receptor for Ybt. In this study, we examined the role of the fyuA gene in Y. pestis virulence using different challenging ways and explored the underlying mechanisms. The BALB/c mouse infection assay showed that the virulence of the mutant strains (ΔfyuA and ΔfyuAGCAdel) was lower when compared with that of the wild-type (WT) strain 201. Furthermore, the attenuation of virulence of the mutant strains via subcutaneous and intraperitoneal challenges was far greater than that via intravenous injection. Iron supplementation restored lethality during subcutaneous challenge with the two mutants. Thus, we speculated that the attenuated virulence of the mutant strains toward the mice may be caused by dysfunctional iron uptake. Moreover, ΔfyuA and ΔfyuAGCAdel strains exhibited lower survival rates in murine RAW264.7 macrophages, which might be another reason for the attenuation. We further explored the transcriptomic differences between the WT and mutant strains at different temperatures and found that the expressions of genes related to Ybt synthesis and its regulation were significantly downregulated in the mutant strains. This finding indicates that fyuA might exert a regulatory effect on Ybt. Additionally, the expressions of the components of the type III secretion system were unexpectedly upregulated in the mutants, which is inconsistent with the conventional view that the upregulation of the virulence genes enhances the virulence of the pathogens.

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“…Cell viability test reveals PSC, PCZ, In order to further compare the cell protective effect of PSCZ, we additionally used an xCELLigence (RTCA) instrument to monitor the survival and growth of these cells in a quantitative, real-time, dynamic manner. This assay approach is superior to traditional endpoint-based viability assays that assess membrane permeability (Yah and Simate, 2020), enabling the assessment of cell survival in a label-free and automated manner throughout a given experimental period (Ceti_n and Topcul, 2019). RTCA approach was used to measure the survival and growth of cells over a 24 h period following injury with H 2 O 2 and these different nanoenzymes preparations.…”

Section: Assessment Of the Ability Of Pscz To Scavenge Reactive Oxygen Species In Vitromentioning

confidence: 99%

Treatment of Acute Kidney Injury Using a Dual Enzyme Embedded Zeolitic Imidazolate Frameworks Cascade That Catalyzes In Vivo Reactive Oxygen Species Scavenging

Hou

Shi

Zhang

et al. 2022

Front. Bioeng. Biotechnol.

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Significant advances have been made in recent years for the utilization of natural enzymes with antioxidant properties to treat acute kidney injury (AKI). However, these enzymes have been of limited clinical utility because of their limited cellular uptake, poor pharmacokinetic properties, and suboptimal stability. We employed a novel biomimetic mineralization approach to encapsulate catalase (CAT) and superoxide dismutase (SOD) in a zeolitic imidazolate framework-8 (ZIF-8). Next, this SOD@CAT@ZIF-8 complex was anchored with MPEG2000-COOH to yield an MPEG2000-SOD@CAT@ZIF-8 (PSCZ) composite. The composite was then used as a stable tool with antioxidant properties for the integrated cascade-based treatment of AKI, remarkably improved intracellular enzyme delivery. This dual-enzyme-embedded metal-organic framework could effectively scavenge reactive oxygen species. In conclusion, the ZIF-8-based “armor plating” represents an effective means of shielding enzymes with improved therapeutic utility to guide the precision medicine-based treatment of AKI.

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Evaluation of the cytotoxic effect of Ly2109761 on HeLa cells using the xCELLigence RTCA system

Cited by 10 publications

References 18 publications

Isoimperatorin Induces Apoptosis of Nasopharyngeal Carcinoma Cells via the MAPK/ERK1/2 Signaling Pathway

Isoimperatorin Induces Apoptosis of Nasopharyngeal Carcinoma Cells via the MAPK/ERK1/2 Signaling Pathway

Attenuation of Yersinia pestis fyuA Mutants Caused by Iron Uptake Inhibition and Decreased Survivability in Macrophages

Treatment of Acute Kidney Injury Using a Dual Enzyme Embedded Zeolitic Imidazolate Frameworks Cascade That Catalyzes In Vivo Reactive Oxygen Species Scavenging

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