Mulberry Anthocyanins Induce Leukemia WEHI-3 Cells Apoptosis, Autophagy, Differentiation and prolong leukemic mice survival

Abstract

Recently, interest in dietary phytochemicals and Chinese herbs for potential cancer chemoprevention has increased substantially owing to their multiple capacities of a combination of several distinct intracellular events (such as cell proliferation, apoptosis, angiogenesis, and metastasis). In our previous studies, we showed that Mulberry anthocyanins (MACs), a natural polyphenol product extracted from Morus, have the antitumor effects on various cancers, such as gastric cancer, melanoma, and hepatoma. Acute myeloid leukemia (AML) patients usually suffer from the side effects of chemotherapy. We investigated effects of MACs on murine Leukemia WEHI-3 cells in vitro. Firstly, 50% of WEHI-3 cells were decreased after treatment with 1.76 mg/ml and 1.48 mg/ml for 24 and 48 hr by MTT assay, respectively. The results of flow cytometry assay showed that low does MACs caused the WEHI3 cells apoptosis when they were exposed to high dose MACs. In addition, caspase-9 was activated from pro-caspase to cleaved-caspase in response to treatment MACs with WEHI-3 cells. Immunoflurescence analysis and western blotting assay demonstrated that MACs caused WEHI3 cells death by triggering autophagic flux. By co-treatment with ATRA, MACs also increased CD11b and GM-CSF level, decreased M-CSF expression, which led to WEHI3 cells differentiation. Moreover, co-treated ATRA and MACs cells showed the suppression of MEK, ERK and STAT1 phosphorylation. Altogether, MACs can prevent the growth of WEHI-3 cells for potentially combination chemotherapy therapy to prevent side effects of acute myeloid leukemia.