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Evaluation of antitumoral effect of mistletoe fruit extract on Ehrlich ascites tumor cells with muse cell analyzer and argyrophilic nucleolar organizer region staining method

Şükrü Ateş

Şükrü Ateş

Department of Anatomy, Faculty of Medicine, Yozgat Bozok UniversityYozgat, Turkey
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Ateş, Şükrü
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Harun Ülger

Harun Ülger

Department of Anatomy, Faculty of Medicine, Erciyes UniversityKayseri, Turkey
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Ülger, Harun
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Seher Yilmaz

Seher Yilmaz

Department of Anatomy, Faculty of Medicine, Yozgat Bozok UniversityYozgat, Turkey
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Yilmaz, Seher
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Gökçe Şeker Karatoprak

Gökçe Şeker Karatoprak

Department of Pharmacognosy, Faculty of Pharmacy, Erciyes UniversityKayseri, Turkey
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Karatoprak, Gökçe Şeker
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Özge Al

Özge Al

Department of Anatomy, Faculty of Medicine, Erciyes UniversityKayseri, Turkey
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Al, Özge
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Sümeyye Uçar

Sümeyye Uçar

Department of Anatomy, Faculty of Medicine, Erciyes UniversityKayseri, Turkey
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Uçar, Sümeyye
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Mustafa Taştan

Mustafa Taştan

Department of Anatomy, Faculty of Medicine, Lokman Hekim UniversityAnkara, Turkey
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Taştan, Mustafa
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Adem Tokpinar

Adem Tokpinar

Department of Anatomy, Faculty of Medicine, Yozgat Bozok UniversityYozgat, Turkey
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Tokpinar, Adem
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Şerife Alpa

Şerife Alpa

Department of Anatomy, Faculty of Medicine, KTO Karatay UniversityKonya, Turkey
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Ammad Ahmad Farooqi

Ammad Ahmad Farooqi

Institute of Biomedical and Genetic Engineering (IBGE)Islamabad, Pakistan
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Farooqi, Ammad Ahmad
  
Jul 11, 2022
Evaluation of antitumoral effect of mistletoe fruit extract on Ehrlich ascites tumor cells with muse cell analyzer and argyrophilic nucleolar organizer region staining method's Cover Image
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Article Category: Original Study
Published Online: Jul 11, 2022
Page range: 209 - 219
Received: Mar 05, 2021
Accepted: Jan 04, 2022
DOI: https://doi.org/10.2478/ahem-2022-0014
Keywords
© 2022 Şükrü Ateş et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1

Experimental processes in this study
Experimental processes in this study

Fig. 2

The raw graphics taken from the Muse® Cell Analyzer. A: Muse® Annexin V and Dead Cell Assay, B: Muse® Cell Cycle Assay, C: Muse® Mitopotential Assay
The raw graphics taken from the Muse® Cell Analyzer. A: Muse® Annexin V and Dead Cell Assay, B: Muse® Cell Cycle Assay, C: Muse® Mitopotential Assay

Fig. 3

Data of the annexin V & dead cell assay at the end of 24, 48, and 72 hours of incubation. p<0.05 was considered statistically significant. Data are expressed as mean ± SEM (standard error of mean). There is no statistically significant difference between the groups containing the same letter (p> 0.05). A: Live cell count (%), B: Early apoptotic cell count (%), C: Late apoptotic cell count (%), D: Total apoptotic cell count (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups
Data of the annexin V & dead cell assay at the end of 24, 48, and 72 hours of incubation. p<0.05 was considered statistically significant. Data are expressed as mean ± SEM (standard error of mean). There is no statistically significant difference between the groups containing the same letter (p> 0.05). A: Live cell count (%), B: Early apoptotic cell count (%), C: Late apoptotic cell count (%), D: Total apoptotic cell count (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups

Fig. 4

Data of the cell cycle assay at the end of 24, 48, and 72 hours of incubation. p <0.05 was considered statistically significant. Data are expressed as mean ± SEM. There is no statistically significant difference between groups containing the same letter (p> 0.05). A: Cell cycle distribution-G0 / G1 stage (%), B: Cell cycle distribution-S stage (%), C: Cell cycle distribution-G2 / M stage (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups
Data of the cell cycle assay at the end of 24, 48, and 72 hours of incubation. p <0.05 was considered statistically significant. Data are expressed as mean ± SEM. There is no statistically significant difference between groups containing the same letter (p> 0.05). A: Cell cycle distribution-G0 / G1 stage (%), B: Cell cycle distribution-S stage (%), C: Cell cycle distribution-G2 / M stage (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups

Fig. 5

Data of the mitopotential assay at the end of 24, 48, and 72 hours of incubation. p <0.05 was considered statistically significant. Data are expressed as mean ± SEM. There is no statistically significant difference between the groups containing the same letter (p> 0.05). A: Live depolarized cell count (%), B: Dead depolarized cell count (%), C: Total depolarized cell count (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups
Data of the mitopotential assay at the end of 24, 48, and 72 hours of incubation. p <0.05 was considered statistically significant. Data are expressed as mean ± SEM. There is no statistically significant difference between the groups containing the same letter (p> 0.05). A: Live depolarized cell count (%), B: Dead depolarized cell count (%), C: Total depolarized cell count (%). 5-FU: 5-fluorouracil; 100, 200, 400, 800 μg/ml: Mistletoe groups

Fig. 6

Image examples of the AgNOR staining cells. A. Control group, B. 5-FU group, C. 100 μg / ml mistletoe group, D. 200 μg/ml mistletoe group, E. 400 μg / ml mistletoe group, F. 800 μg/ml mistletoe group
Image examples of the AgNOR staining cells. A. Control group, B. 5-FU group, C. 100 μg / ml mistletoe group, D. 200 μg/ml mistletoe group, E. 400 μg / ml mistletoe group, F. 800 μg/ml mistletoe group

Mean AgNOR number after 24, 48, and 72 hours of incubation

Hours/Groups Control 5-FU 100 μg/ml 200 μg/ml 400 μg/ml 800 μg/ml p
24 hours 3.54±1.26a 2.82±1.06b 2.98±1.16ab 1.74±0.77c 1.74±1.00c 1.80±1.19c <0.001
48 hours 3.68±1.55a 3.16±1.07a 2.40±0.78b 2.12±1.25b 1.96±0.75b 1.28±0.53c <0.001
72 hours 1.52±0.64a 2.20±1.08bc 2.26±1.24c 2.20±1.01bc 1.68±0.76ab 1.74±0.85abc <0.001

Total phenolic and flavonoid contents of mistletoe fruit

Total phenol (mgGAE/gextract) Total flavonoid (mgCAE/gextract)
Mistletoe fruit 6.24±2.26 4.82±1.36

TAA / NA value at the end of 24, 48, and 72 hours of incubation

Hours/Groups Control 5-FU 100 μg/ml 200 μg/ml 400 μg/ml 800 μg/ml p
24 hours 0.11±0.04ab 0.14±0.04b 0.10±0.03ac 0.13±0.06ab 0.08±0.04c 0.10±0.05ac <0.001
48 hours 0.27±0.15a 0.23±0.05a 0.17±0.08b 0.13±0.12bc 0.09±0.05c 0.09±0.07c <0.001
72 hours 0.10±0.04ac 0.12±0.04ab 0.13±0.05b 0.10±0.06ac 0.08±0.04c 0.11±0.04bc <0.001
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