Naoki Nanashima1, Kayo Horie2, Toshiko Tomisawa3, Mitsuru Chiba1, Manabu Nakano1, Toshifumi Fujita4, Hayato Maeda5, Maiko Kitajima2, Shizuka Takamagi4, Daishi Uchiyama6, Jun Watanabe2, Toshiya Nakamura1, Yoji Kato7. 1. Department of Biomedical Sciences, Hirosaki University Graduate School of Health Sciences, Hirosaki, Japan. 2. Department of Pathologic Analysis, Hirosaki University Graduate School of Health Sciences, Hirosaki, Japan. 3. Department of Health Promotion, Hirosaki University Graduate School of Health Sciences, Hirosaki, Japan. 4. Department of Disability and Health, Hirosaki University Graduate School of Health Sciences, Hirosaki, Japan. 5. Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki, Japan. 6. Center for Joint Research, Hirosaki University, Hirosaki, Japan. 7. Faculty of Education, Hirosaki University, Hirosaki, Japan.
Abstract
SCOPE: Blackcurrants (Ribes nigrum L., Grossulariaceae) contain high amounts of anthocyanin polyphenols, which have antioxidant and anti-carcinogenic health benefits. This study analyzed the potential phytoestrogenic effects of blackcurrant extract (BCE) in breast cancer (MCF-7) and human endometrial cancer (Ishikawa) cell lines that over-express estrogen receptor alpha (ERα), as well as in immature female rats. METHODS AND RESULTS: Microarray analysis and Ingenuity® Pathway Analysis showed that BCE activated the ERα pathway, whereas quantitative-PCR confirmed that BCE and four types of anthocyanins up-regulated genes downstream of ERα. BCE (0.1-1.0 μg/mL) and anthocyanins (0.1-10 μM) induced MCF-7 cell proliferation; however, this effect was blocked by ER antagonist fulvestrant. Flow cytometry showed that anthocyanins reduced and increased the number of MCF-7 cells in the G0/G1 and G2/M phases, respectively. Anthocyanins stimulated ERα transcriptional activity in human ERα reporter assays and induced alkaline phosphatase activity in Ishikawa cells. Competition assays and in silico analysis indicated that anthocyanins bind to ERα. Finally, BCE focally induced stratification of columnar epithelial cells in the rat uterus and increased cytoplasmic mucin levels in these cells. CONCLUSION: These results suggest that blackcurrant anthocyanins act as phytoestrogens in vitro and in vivo.
SCOPE: Blackcurrants (Ribes nigrum L., Grossulariaceae) contain high amounts of anthocyanin polyphenols, which have antioxidant and anti-carcinogenic health benefits. This study analyzed the potential phytoestrogenic effects of blackcurrant extract (BCE) in breast cancer (MCF-7) and humanendometrial cancer (Ishikawa) cell lines that over-express estrogen receptor alpha (ERα), as well as in immature female rats. METHODS AND RESULTS: Microarray analysis and Ingenuity® Pathway Analysis showed that BCE activated the ERα pathway, whereas quantitative-PCR confirmed that BCE and four types of anthocyanins up-regulated genes downstream of ERα. BCE (0.1-1.0 μg/mL) and anthocyanins (0.1-10 μM) induced MCF-7 cell proliferation; however, this effect was blocked by ER antagonist fulvestrant. Flow cytometry showed that anthocyanins reduced and increased the number of MCF-7 cells in the G0/G1 and G2/M phases, respectively. Anthocyanins stimulated ERα transcriptional activity in human ERα reporter assays and induced alkaline phosphatase activity in Ishikawa cells. Competition assays and in silico analysis indicated that anthocyanins bind to ERα. Finally, BCE focally induced stratification of columnar epithelial cells in the rat uterus and increased cytoplasmic mucin levels in these cells. CONCLUSION: These results suggest that blackcurrant anthocyanins act as phytoestrogens in vitro and in vivo.
Authors: Reshmi Akter; Jong Chan Ahn; Jinnatun Nahar; Muhammad Awais; Zelika Mega Ramadhania; Se-Woung Oh; Ji-Hyung Oh; Byoung Man Kong; Esrat Jahan Rupa; Dong Wong Lee; Deok Chun Yang; Se Chan Kang Journal: Front Pharmacol Date: 2022-09-29 Impact factor: 5.988