Abstract
ABSTRACTBackground: Anthocyanins have been proven to affect multiple cancer-associated processes in different cancer cell lines. However, relatively few studies have investigated the effects of blueberry anthocyanins on metastatic melanoma. Thus, this study focuses on evaluating the chemopreventive potential of blueberry anthocyanins and their aglycones (anthocyanidins) in B16-F10 melanoma cells.Methods: Blueberry anthocyanin and anthocyanidin extracts were prepared mainly by combined chromatography techniques. Their antiproliferative and proapoptotic effects on B16-F10 cells were evaluated by MTT assay, calcein acetoxymethyl ester/propidium iodide (calcein-AM/PI) staining, and flow cytometry of the cell cycle and apoptosis.Results: The MTT and calcein-AM/PI staining results showed that both anthocyanin (purity of 62.5%) and anthocyanidin (75.1%) extracts could significantly inhibit the viability and proliferation of B16-F10 cells in a dose-dependent manner, while anthocyanidin extracts exhibited significantly higher (p < 0.05) cytotoxicity than anthocyanin extracts. Furthermore, anthocyanin and anthocyanidin extracts blocked cell cycle procession at the G0/G1 phase below 400 and 200 μg/mL, and induced early apoptosis below 400 and 300 μg/mL, respectively.Conclusions: These data suggest that both anthocyanin and anthocyanidin extracts inhibit the proliferation and trigger the apoptosis of B16-F10 cells, and anthocyanidin extracts may be a more promising candidate in preventing metastatic melanoma than anthocyanin extracts.
Highlights
Metastasis is a complex multistep process by which malignant tumors can spread to distal organs [1]
The chemical structures of anthocyanins and anthocyanidins in the two anthocyan samples were confirmed through the combined interpretation of ultraviolet–visible spectra, predicted molecular polarities, congruent retention time with the available standards, HPLC-DAD–electrospray ionization–tandem mass spectrometry data, and the identities of anthocyans in other blueberry species comparable to those in earlier studies [24]
Following the separation procedures of extraction, partition, column chromatography, and solidphase extraction, the anthocyanin extracts were obtained, and the HPLC profiles in Figure 1(B) indicated that most the 16 monomeric anthocyanins were obtained, despite a slight difference in the ratios of peak areas of each anthocyanin compared with the anthocyanin profiles in blueberry fruits (Figure 1(A))
Summary
Metastasis is a complex multistep process by which malignant tumors can spread to distal organs [1]. Tumor metastasis is regarded as the most common cause of mortality in cancer patients. It is believed that phytochemicals such as polyphenols in fruits or vegetables can induce cell apoptosis or cell cycle arrest in tumorderived cells, while having a relatively low toxicity [4]. This study focuses on evaluating the chemopreventive potential of blueberry anthocyanins and their aglycones (anthocyanidins) in B16-F10 melanoma cells. Methods: Blueberry anthocyanin and anthocyanidin extracts were prepared mainly by combined chromatography techniques. Their antiproliferative and proapoptotic effects on B16-F10 cells were evaluated by MTT assay, calcein acetoxymethyl ester/propidium iodide (calcein-AM/PI) staining, and flow cytometry of the cell cycle and apoptosis
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