Bog Bilberry (Vaccinium uliginosum L.) Extract Reduces Cultured Hep-G2, Caco-2, and 3T3-L1 Cell Viability,Affects Cell Cycle Progression,and Has Variable Effects on Membrane Permeability

ABSTRACT: Bog bilberry (Vaccinium uliginosum L.) is a blue-pigmented edible berry related to bilberry (Vaccinium myrtillus L.) and the common blueberry (Vaccinium corymbosum). The objective of this study was to investigate the effect of a bog bilberry anthocyanin extract (BBAE) on cell growth, membrane permeability, and cell cycle of 2 malignant cancer cell lines, Caco-2 and Hep-G2, and a nonmalignant murine 3T3-L1 cell line. BBAE contained 3 identified anthocyanins. The most abundant anthocyanin was cyanidin-3-glucoside (140.9 ± 2.6 μg/mg of dry weight), followed by malvidin-3-glucoside (10.3 ± 0.3 μg/mg) and malvidin-3-galactoside (8.1 ± 0.4 μg/mg). Hep-G2 LC50 was calculated to be 0.563 ± 0.04 mg/mL, Caco-2 LC50 was 0.390 ± 0.30 mg/mL and 0.214 ± 0.02 mg/mL for 3T3-L1 cells. LDH release, a marker of membrane permeability, was significantly increased in Hep-G2 cells and Caco-2 cells after 48 and 72 h compared to 24 h. The increase was 21% at 48 h and 57% at 72 h in Caco-2 cells and 66% and 139% in Hep-G2 cells compared to 24 h. However, 3T3-L1 cells showed an unexpected significant lower LDH activity (P ≤ 0.05) after 72 h of exposure corresponding to a 21% reduction in LDH release. BBAE treatment increased sub-G1 in all 3 cell lines without influencing cells in the G2/M phase. BBAE treatment reduced the growth and increased the accumulation of sub-G1 cells in 2 malignant and 1 nonmalignant cell line; however, the effect on membrane permeability differs considerably between the malignant and nonmalignant cells and may in part be due to differences in cellular membrane composition.