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From
Characterization of Anti-Cancer Effects of the Ethyl Acetate Fraction from
Orostachys
japonicus
on HT-29 Human Colon Cancer Cells
Hyun Ji Lee, Dong Seok Lee
Journal of Food and Nutrition Research
.
2025
, 13(3), 127-139 doi:10.12691/jfnr-13-3-2
Fig
ure
1
.
Effects of kaempferol alone (a), quercetin alone (b), OJE (c), and kaempferol alone, quercetin alone, OJE, and combinations of kaempferol, quercetin, and/or OJE (d) on viability of RAW 264.7 macrophage cells measured by cytotoxicity assay for 12 and 24 h. The results are presented as the mean ± standard deviation (SD) of three independent experiments. Statistical significance was determined as *p < 0.05, **p < 0.01, and ***p < 0.001
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Fig
ure
2
.
Inhibitory Effects of kaempferol (e), kaempferol + OJE (f), quercetin (g), quercetin + OJE (h), kaempferol + quercetin (i), kaempferol + quercetin + OJE (j), and OJE (k) on the proliferation of HT-29 cells.
Cell viability was assessed using the MTS assay. The results are presented as the mean ± standard deviation (SD) of three independent experiments. Statistical significance was determined as *p < 0.05, **p < 0.01, and ***p < 0.001
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Fig
ure
3
.
Nuclear morphological changes indicative of inducing apoptosis observed in HT-29 cells treated with kaempferol (a), quercetin (b), and kaempferol alone, quercetin alone, a mixture of kaempferol and quercetin, a mixture of kaempferol, quercetin, and OJE, and OJE (c). They were then stained with the DNA-specific fluorochrome DAPI. White arrows signify apoptotic nuclear morphological changes
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Fig
ure
4
.
Flow cytometry analysis of apoptosis progression in HT-29 cells treated with kaempferol alone, quercetin alone, kaempferol + quercetin, kaempferol + quercetin + OJE, and OJE. The cells were exposed to various concentrations (kaempferol 100 µM, quercetin 100 µM, kaempferol 50 µM + quercetin 50 µM, kaempferol 33 µM + quercetin 33 µM + OJE 33µg/mL, and OJE 100µg/mL) fo 12 h. The cells were labeled with monoclonal antibodies specific to Annexin V-FITC and PI. Based on the staining pattern, cells were categorized as follows: Annexin−/PI− (LL) representing viable cells, Annexin+/PI− (LR) indicating cells undergoing apoptosis, and Annexin+/PI+ (UR) representing cells in end-stage of apoptosis or already deceased
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Fig
ure
5
.
Flow cytometry analysis of cell cycle arrest in HT-29 cells treated with kaempferol (a), quercetin (b), kaempferol alone, quercetin alone, K + Q (kaempferol + quercetin), K + Q + O (kaempferol + quercetin + OJE), and OJE (c). Histograms represent sub-G1, G1, S, and G2/M phases of HT-29 cells. The results were expressed as percentage of total treated cells. Three experiments showed similar results
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Fig
ure
6
.
Wound healing assay for measurement of inhibitory effects of OJE, kaempferol, and quercetin alone or in combination on the migration of HT-29 cells. Three experiments showed similar results
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Fig
ure
7
.
Western blot analysis of apoptosis-related key biomarkers produced or remained in HT-29 cells treated with kaempferol (a), quercetin (b), and single samples or combinations of kaempferol, quercetin, and/or OJE (c). The density of bands was quantitated. GAPDH was used as an internal control. Band intensities were measured by densitometry in three separate experiments with similar results
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Fig
ure
8.
Western blot analysis of cell cycle arrest-related key biomarkers produced or remained in HT-29 cells treated with quercetin (a), and single samples or combinations of kaempferol, quercetin, and/or OJE (b). The density of bands was quantitated. GAPDH was used as an internal control. Band intensities were measured by densitometry in three separate experiments with similar results
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Fig
ure
9
.
Western blot analysis of anti-metastasis-related key biomarkers produced or remained in HT-29 cells treated with quercetin (a), and single samples or combinations of kaempferol, quercetin, and/or OJE (b). The density of bands was quantitated. GAPDH was used as an internal control. Band intensities were measured by densitometry in three separate experiments with similar results
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Fig
ure
10
.
Western blot analysis of upstream signal transduction pathways-related key biomarkers produced or remained in HT-29 cells treated with single samples or combinations of kaempferol, quercetin, and/or OJE. The density of bands was quantitated. GAPDH was used as an internal control. Band intensities were measured by densitometry in three separate experiments with similar results
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