Abstract
Green tea consumption is associated with chemoprevention of many cancer types. Fresh tea leaves are rich in polyphenolic catechins, which can constitute up to 30% of the dry leaf weight. While the polyphenols of green tea have been well investigated, it is still largely unknown, whether or not non-phenolic constituents also reveal chemopreventive and anti-metastatic effects. In this study, we investigated the effects of a fraction of green tea rich in phenolic compounds (PF), a non-phenolic fraction (NPF), which contains glyceroglycolipids (GGL), and a pure glyceroglycolipid compound isolated from the non-phenolic fraction in human cancer. Dried green tea leaves were extracted and applied to a Sephadex LH-20 column. The resazurin reduction assay was used to investigate the cytotoxicity of green tea samples toward human HepG2 hepatocellular carcinoma and normal AML12 hepatocytes cells. Gene expression profiling was performed by mRNA microarray hybridization and the microarray results were validated by RT-PCR. The scratch migration assay was used to investigate the effects of green tea samples on cell migration in vitro. The changes of microtubule dynamics were observed using fluorescence microscopy. PF and NPF were prepared from methanol extract of green tea. A GGL was isolated from NPF. All three green tea samples did not show significant cytotoxic activity up to 10 μg/mL in both HepG2 and AML12 cells, whereas cytotoxicity of the control drug doxorubicin was observed with both cell lines (IC50 on AML12: 0.024 μg/mL, IC50 on HepG2: 2.103 μg/mL). We identified three sets of genes differentially expressed upon treatment with the green tea samples. The genes were associated with cytoskeleton formation, cellular movement, and morphology. The correlation coefficients between mRNA expression values determined by microarray and RT-PCR were R = 0.94. HepG2 and U2OS cells treated with green tea extracts showed the delayed closures. Besides, the number of distinct tubulin filaments decreased upon treatment with green tea samples. We identified not only PF, but also glyceroglycolipids in NPF as contributing factors to the chemopreventive effects of green tea. Both PF and NPF of green tea inhibited cancer cell migration by the disassembly of microtubules, even though they were not cytotoxic.
Highlights
Tea is one of the most popular beverages around the world
Our study focused on investigating the efficacy of a fraction of green tea rich in phenolic compounds, a non-phenolic fraction, which contains glyceroglycolipids, and a pure compound belonging to glyceroglycolipid class, which was isolated from the non-phenolic fraction
The aim of this study was to investigate the effects of green tea constituents in human cancer cells
Summary
Tea is one of the most popular beverages around the world. It is obtained from the leaves of Camellia sinensis L. Fresh tea leaves are rich in polyphenols known as catechins, which may constitute up to 30% of the dry leaf weight (Cabrera et al, 2006; Chacko et al, 2010). Other polyphenols include flavonols and their glycosides and one compound unique to tea, theogallin (3-galloylquinic acid) (Saleh et al, 2013). Green tea leaves contain three main classes of compounds, which are known to affect human health, i.e., xanthic bases (caffeine and theophylline), essential oils, and polyphenolic compounds (Graham, 1992). The nonphenolic components of green tea have not been explored in detail and their biological effects are unknown
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