Elsevier

Phytomedicine

Volume 21, Issue 12, 15 October 2014, Pages 1746-1752
Phytomedicine

Hispolon from Phellinus linteus possesses mediate caspases activation and induces human nasopharyngeal carcinomas cells apoptosis through ERK1/2, JNK1/2 and p38 MAPK pathway

https://doi.org/10.1016/j.phymed.2014.07.013Get rights and content

Abstract

Hispolon, a phenol compound isolated from Phellinus linteus (PL), possesses anti-inflammatory, antiproliferative, and antioxidant effects. However, the effects of hispolon on human nasopharyngeal carcinomas have yet to be evaluated. Here, the molecular mechanism by which hispolon anticancer effects in human nasopharyngeal carcinomas cells was investigated. The results showed that hispolon significantly inhibited cell proliferation of HONE-1 and NP-039 cell lines. Furthermore, hispolon induced apoptosis through caspases-3, -8, and -9 activations and PARP cleavage in dose- and time-dependent manner in HONE-1 and NP-039 cells. Moreover, hispolon also showed that increase phosphorylation of ERK1/2, p38 MAPK and JNK1/2 in dose- and time-dependent manner by western blot analysis. However, hispolon-induced activation of the caspase-3, -8 and -9 significantly abolished by inhibition of p38 MAPK and JNK1/2 specific inhibitors. In this study, we determine that the effects of hispolon on the apoptosis and related regulation mechanism in HONE-1 and NPC-039 cells takes place. Our findings revealed that hispolon may be a useful candidate as a chemotherapeutic agent for NPC therapy.

Introduction

Phellinus linteus (Berkeley & Curtis), a famous medical fungus of the genus Phellinus, has been used as a traditional medicinal mushroom, is commonly called “Sanghwang” in Taiwan. It is popular in oriental countries such as China, Korea, and Japan for many years to treat various diseases, such as gastroenteric disorders, inflammation, tumors and lymphatic diseases (Chou et al., 2013, Lim et al., 2004). Various bioactive components, such as polysaccharides, proteoglycans, furan derivatives, hispidin and hispolon, have been identified from P. linteus (Kim et al., 2004, Kim et al., 2006, Zhu et al., 2008). Previous studies show that PL suppresses cellular proliferation and induces apoptosis in lung and prostate cancer cells (Guo et al., 2007, Zhu et al., 2007). The antimetastatic effects of PL were demonstrated against in vivo colon cancer and melanoma models (Lee et al., 2005).

Hispolon, a phenol compound isolated from PL, has been reported that the effect of anti-oxidant, anti-inflammatory, anti-proliferative, and exerts protective effects on acute liver damage (Ali et al., 1996, Huang et al., 2012). Additionally, hispolon also confirms that an antitumor effect in various cancer cell lines takes place (Chen et al., 2008, Huang et al., 2011, Lu et al., 2009). However, there are no reports concerning the anticancer effects of hispolon on human nasopharyngeal carcinomas.

Nasopharyngeal carcinoma (NPC) is the most common cancer originating in the nasopharynx, where the nasal passages and auditory tubes join the remainder of the upper respiratory tract. NPC occurs in children and adults. Despite occurring commonly in Southeast Asia and southern provinces of China, it rarely occurs in Northern China, Europe, and America (Cao et al., 2011, Chou et al., 2008). It is a distinct entity of head and neck cancers because of its characteristic epidemiology, pathogenesis, and association with the Epstein–Barr virus (Wei and Sham, 2005). Importantly, It is a metastasis of cancer cells to the neck lymph nodes, which can occur in up to 75% of NPC patients, which represents an adverse prognostic factor of the disease (Chua et al., 2001). In the present study, we investigated the cytotoxic effects of hispolon on human NPC cell lines, HONE-1 and NPC-039, and its underlying mechanisms in vitro.

Section snippets

Chemicals

Hispolon, ≥98% (HPLC), powder was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Stock solution of hispolon was made at 12.5, 25, 50 and 100 μM concentration in DMSO and stored at −20 °C. The final concentration of DMSO for all treatments was consistently less than 0.1%. 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) was obtained from Sigma Chemical Co. (St. Louis, MO, USA). Specific inhibitors for ERK1/2 (U0126), JNK1/2 (SP600125) or p38 (SB202190) were purchased

The effects of cell viability in hispolon-treated human nasopharyngeal carcinoma cell lines

The chemical structure of hispolon was shown in Fig. 1A. To determine the cytotoxicity and the cell viability of hispolon on human NPC cell lines, cells were treated with different concentrations of hispolon (0–100 μM). The results show that hispolon significantly inhibited cell viability in a dose-dependent and time-dependent manner by MTT assay (Fig. 1B and C). We further studied the anti-proliferation activity of hispolon on human NPC cell lines by cell counting. Fig. 1D shows that hispolon

Discussion

Nasopharyngeal carcinoma (NPC) is a rare tumor arising from the epithelium of the nasopharynx. Natural herbal products have a promising and potential role in developing novel chemotherapeutics in the prevention and/or treatment of several chronic diseases (Chen et al., 2006, Chen et al., 2008, Lu et al., 2009). Chemoprevention is an active cancer preventive strategy to suppress, delay, or reverse human carcinogenesis. In the present study, we investigated the apoptosis of human nasopharyngeal

Competing interest

The authors declare that there are no conflicts of interest.

Acknowledgements

This study was supported by grants from Ministry of Science and Technology, Taiwan (MOST 103-2314-B-371-007-MY2) and Changhua Christian Hospital (103-CCH-IRP-071). The authors of the manuscript do not have a direct financial relation with the commercial identity mentioned in this paper. This study will be presented in the MedicReS 4th world congress on “good medıcal research” to be held in New York City, New York, USA, from October 16 to 18, 2014.

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