The comparative study among different fractions of muscadine grape ‘Noble’ pomace extracts regarding anti-oxidative activities, cell cycle arrest and apoptosis in breast cancer

Figures & data

Table 1. 80% and 50% maximum inhibitory concentrations of different fractions.

Group	Concentration (µg DW/mL)
	IC50 (low concentration)	IC80 (high concentration)
F1	17.82 ± 2.42^a	29.64 ± 1.50^a
F2	9.75 ± 0.52^b	20.85 ± 2.37^b
F3	12.88 ± 2.69^ab	25.00 ± 1.73^ab

F1: mixture of flavonoids, phenolic acids and EA; F2: mixture of tannins; F3: mixture of anthocyanidins and EA. The same letter in the same column indicates no significant differences (p ≥ 0.05).

Figure 1. Inhibition rates for MDA-MB-231 cells after treatment with isolated fractions of ‘Noble’ pomace. F1: mixture of flavonoids, phenolic acids and EA; F2: mixture of tannins; F3: mixture of anthocyanidins and EA.

Table 2. Anti-oxidative activities of different fractions at low and high concentrations.

	DPPH (µM TE/mL)		ABTS (µM TE/mL)		FRAP (µM TE/mL)
	Low conc.	High conc.	Low conc.	High conc.	Low conc.	High conc.
F1	1.34 ± 0.20^b	2.27 ± 0.34^b	2.33 ± 0.25^a	3.95 ± 0.42^a	2.37 ± 0.21^b	4.01 ± 0.36^b
F2	1.24 ± 0.31^b	2.63 ± 0.65^b	1.91 ± 0.42^a	4.06 ± 0.90^a	2.16 ± 0.06^b	4.60 ± 0.13^b
F3	2.23 ± 0.18^a	4.44 ± 0.38^a	2.57 ± 0.14^a	5.13 ± 0.27^a	2.94 ± 0.05^a	5.87 ± 0.09^a

Low conc.: low concentration; High conc.: high concentration. F1: mixture of flavonoids, phenolic acids and EA; F2: mixture of tannins; F3: mixture of anthocyanidins and EA. The same letter in the same column indicates no significant differences (p ≥ 0.05).

Figure 2. Plots from the cell cycle arrest assay using different fractions at low and high concentrations. Control: solvent treatment; F1 (F2, F3)_L: low concentrations of F1 (F2, F3); F1 (F2, F3)_H: high concentrations of F1 (F2, F3).

Table 3. Proportion of cells in different cell cycle phases after treatment with different fractions at high and low concentration (mean ± standard deviation).

Phase	Concentration	Group
		Control	F1	F2	F3
G0/G1	Low	67.17 ± 0.52^a	59.50 ± 0.21^b	58.49 ± 0.13^c	60.25 ± 0.13^b
	High	67.17 ± 0.52^a	56.90 ± 0.34^b	54.21 ± 1.50^c	55.20 ± 0.06^bc
S	Low	18.22 ± 0.07^c	30.28 ± 0.22^b	34.13 ± 0.38^a	33.16 ± 0.71^a
	High	18.22 ± 0.07^c	36.88 ± 0.21^b	40.28 ± 0.80^a	39.66 ± 0.13^ab
G2/M	Low	14.61 ± 0.48^a	10.22 ± 0.64^b	7.38 ± 0.31^c	6.58 ± 0.22^c
	High	14.61 ± 0.48^a	6.32 ± 0.04^b	5.52 ± 0.63^b	5.15 ± 0.17^b

F1: mixture of flavonoids, phenolic acids and EA; F2: mixture of tannins; F3: mixture of anthocyanidins and EA. The same letter in the same row indicates no significant differences (p ≥ 0.05).

Figure 3. Expression of cell cycle arrest proteins following treatment with different fractions at high or low concentrations. (a) Bands obtained from Western blot analysis; (b) bar charts of the mean integrated density of each band over the corresponding integrated density of GAPDH (control value adjusted to 1). ‘a, b, c’: the same letter above the bars indicates no significant differences (p ≥ 0.05). Control: solvent treatment; F1 (F2, F3)_L: low concentrations of F1 (F2, F3); F1 (F2, F3)_H: high concentrations of F1 (F2, F3).

Table 4. Proportions of normal and apoptotic cells after treatment with different fractions at high and low concentrations (mean ± standard deviation).

Cell	Concentration	Group
		Control	F1	F2	F3
Normal	Low	99.24 ± 0.17^a	89.85 ± 0.45^c	93.45 ± 0.32^b	87.91 ± 0.13^d
	High	99.24 ± 0.17^a	79.17 ± 0.11^c	87.15 ± 0.11^b	77.14 ± 0.16^d
Apoptotic	Low	0.40 ± 0.08^d	10.03 ± 0.37^b	6.47 ± 0.63^c	12.03 ± 0.63^a
	High	0.40 ± 0.08^d	19.63 ± 0.48^b	11.99 ± 0.50^c	21.52 ± 0.58^a

F1: mixture of flavonoids, phenolic acids and EA; F2: mixture of tannins; F3: mixture of anthocyanidins and EA. The same letter in the same row indicates no significant differences (p ≥ 0.05).

Figure 4. Plots from the cell apoptosis assay for different fractions at low and high concentrations. Control: solvent treatment; F1 (F2, F3)_L: low concentrations of F1 (F2, F3); F1 (F2, F3)_H: high concentrations of F1 (F2, F3).

Figure 5. Contents of apoptosis proteins following treatment with different fractions at high or low concentrations. (a) Bands obtained from Western blotting; (b) bar charts of the mean integrated density of each band over the corresponding integrated density of GAPDH (control value adjusted to 1). ‘a, b, c’: the same letter above the bars indicates no significant differences (p ≥ 0.05). Control: solvent treatment; F1 (F2, F3)_L: low concentrations of F1 (F2, F3); F1 (F2, F3)_H: high concentrations of F1 (F2, F3).