The effect of resveratrol, its naturally occurring derivatives and tannic acid on the induction of cell cycle arrest and apoptosis in rat C6 and human T98G glioma cell lines
Introduction
Malignant gliomas are the most common central nervous system tumors and carry the worst clinical prognosis in both adults and children. Although current therapeutic protocols comprise a combination of surgical operation with irradiation and adjuvant chemotherapy, the prognosis for malignant gliomas remains very poor due to their highly aggressive biological behavior and frequent recurrence rate (Gagliano et al., 2010, Van Meir et al., 2010). Thus, the development of new agents able to reactivate cell cycle or cell death programs, particularly apoptosis, is important for the introduction of new therapeutic and/or chemopreventive strategies for malignant gliomas.
Resveratrol (3,5,4′-trihydroxy-trans-stilbene), a naturally occurring polyphenol, is assumed to possess cancer-preventive and cancer-therapeutic properties (Kuršvietienė et al., 2016). Resveratrol suppresses the proliferation of a variety of human cancer cells in vitro, including glioma cells (Gagliano et al., 2010).
In C6 rat glioma cells, resveratrol has been shown to inhibit cell growth and induce apoptosis by increasing caspase-3 mRNA level and enzyme activation (Zhang et al., 2007). However, Michels et al. (2006) demonstrated that resveratrol is not metabolized in C6 rat glioma cells, and accumulates to concentrations that drive the cell to necrosis. Induction of apoptosis by resveratrol has also been reported in human glioma U251, U87 and T98G cells (Jiang et al., 2005, Lin et al., 2011). Metabolism plays an important role in resveratrol anticancer activity.
While resveratrol is the most active, its sulfo- and glucuronide conjugates are quickly excreted and are much less effective (Sun et al., 2013, Wang et al., 2005).
In view of the limited bioavailability of resveratrol, its naturally occurring and synthetic analogs are the subjects of intensive research and development. In this regard, naturally occurring methoxylated stilbenes, such as 3,5-dimethoxy-4′-hydroxy-trans-stilbene (pterostilbene) and 3,5,4′-trimethoxy-trans-stilbene, have been shown to be more efficient in eliciting chemopreventive and/or therapeutic effects than resveratrol (Kapetanovic et al., 2011, Lin et al., 2009, Lin and Ho, 2009, Aldawsari and Velázquez-Martínez, 2015).
Tannic acid, a water-soluble tannin, is, similarly to resveratrol, a common ingredient of grapes and others berry fruits. Thus, their synergic action may be expected.
Our earlier studies have shown that although both polyphenols reduced binding of carcinogenic aromatic hydrocarbons to DNA, tannic acid was a much more potent inhibitor of specific DNA adduct formation (Ignatowicz et al., 2003, Baer-Dubowska and Szaefer, 2013). Moreover, neutrophil modulation of ROS production ultimately leading to cell apoptosis has also been shown in humans (Zielińska-Przyjemska et al., 2015).
While the role of neutrophils infiltrating tumors has been neglected for some time, a significant amount of evidence has recently been gathered on their importance in experimental models as well as in human cancers (Moses and Brandau, 2016). Fossati et al. (1999) showed that most human glioma samples analyzed had significant neutrophil infiltration. Thus, compounds such as polyphenols may target both cancer tissue and infiltrated neutrophils.
In the present study, we compared the ability of resveratrol and its two natural derivatives (pterostilbene and 3,5,4′-trimethoxystilbene) and tannic acid to induce growth inhibition and apoptosis in rat C6 and human T98G glioma cells. An attempt to explain the mechanism of pro-apoptotic activity of these compounds was also undertaken.
Section snippets
Chemicals
Table 1 shows the chemical structure of the examined polyphenols. Resveratrol, tannic acid (purity GC, TLC ≥ 99%), camptothecin, antibiotic solution (10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL), dimethyl sulfoxide (DMSO), Dulbecco's Modified Eagle's Medium (DMEM), Eagle's Minimum Essential Medium (EMEM), fetal bovine serum (FBS), glutamine, propidium iodide (PI), ribonuclease A (RNase A) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT),
The effect of resveratrol, its analogs and tannic acid on C6 and T98G cell viability
Viability of rat C6 and human T98G glioma cells were evaluated using MTT assays. The IC50 values for resveratrol, pterostilbene and tannic acid did not differ significantly, except resveratrol and tannic acid in T98G cells which in general were less susceptible to cytotoxic effect of these polyphenols (Table 1). TMS significantly reduced the C6 glioma cell viability at concentrations ~ 40–50% lower than the other compounds. Based on the results of viability assays, the concentrations of the
Discussion
The non-toxicity and ability of resveratrol to cross the blood-brain barrier (Wang et al., 2002, Xue et al., 2016) suggests the potential usefulness of this agent in the management of brain malignancy.
The rat C6 glioma cell line is considered to be a valuable experimental model system for the study of glioblastoma growth, invasion and metastasis as well as screening for future drug targets and development of novel therapies (Grobben et al., 2002). In this study, we compared the results obtained
Declaration of interest
The authors report no declarations of interests.
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