Elsevier

Biomedicine & Pharmacotherapy

Volume 102, June 2018, Pages 502-509
Biomedicine & Pharmacotherapy

Osthole inhibits the PI3K/AKT signaling pathway via activation of PTEN and induces cell cycle arrest and apoptosis in esophageal squamous cell carcinoma

https://doi.org/10.1016/j.biopha.2018.03.106Get rights and content

Abstract

Esophageal squamous cell carcinoma (ESCC) is one of the most common lethal tumors and is known to be lack of effective therapy. Thus, novel therapeutic strategies are greatly needed for treatment of ESCC. Osthole, a natural active extract, has been documented to have anti-tumor activity. However, the effect of osthole on ESCC cells has not been elucidated. In this study, we demonstrated that osthole could inhibit the ESCC cell proliferation in dose- and time-dependent manner. Osthole treatment also induced G2/M phase arrest and apoptosis of ESCC cells. Furthermore, upon exposure to osthole, the expression of Cyclin B1, Cdc2, Bcl-2, PARP1 and Survivin was decreased, while the expression of BAX, cleaved PARP1, cleaved Caspase3 and cleaved Caspase9 was increased. In addition, osthole treatment elicited upregulation of PTEN and downregulation of PI3K and phosphorylated AKT (p-AKT). Taken together, our study demonstrates that osthole could suppress ESCC proliferation through inducing cell cycle arrest and apoptosis. Moreover, PTEN-PI3K/AKT signaling pathway can be regulated by osthole. Our results indicate that osthole may find therapeutic application in the treatment of ESCC patients.

Introduction

Esophageal cancer is the third most prevalent cancer and the fourth leading cause of cancer death in China, with esophageal squamous cell carcinoma (ESCC) accounting for over 90% of the cases [1,2]. Despite advances in diagnosis and therapy of patients with esophageal cancer, overall five-year survival rate only ranges from 15% to 25% [3]. In order to ameliorate the survival rate, efforts have been made to explore novel anticancer agents, and much attention has been focused on traditional Chinese herbal medicines, due to their wide variety of biological activities as well as low levels of side effects and toxicity [[4], [5], [6]].

Osthole (chemical formula C15H16O3) is an active component extracted from the fruit of Fructus Cnidii, which has long been applied in traditional Chinese medicine to cure impotence, lumbar pain and rheumatic pain [7,8]. In previous reports, osthole has been demonstrated to possess vasorelaxant [9], anti-osteoporotic [10], anti-inflammatory properties [11,12], and to exert anti-allergic [13] and anti-seizure effects [14]. Recently, increasing evidence suggests that osthole has antitumor activity by inducing apoptosis and inhibiting cancer cell growth and metastasis [[15], [16], [17]]. However, the effect of osthole on human ESCC cells and its underlying mechanism have not been reported yet.

In this study, we demonstrated that osthole remarkably inhibited ESCC cell proliferation in dose-and time-dependent manner, and induce G2/M phase arrest and apoptosis of ESCC cells. Moreover, PTEN-PI3K/AKT signaling pathway can be modulated by osthole treatment. Thus, osthole might be a novel active component to exploit an effective medicine for treatment of ESCC patients.

Section snippets

Reagents, antibodies and kits

Osthole (>98%) powder was obtained from Chengdu Must Bio-Technology Co., Ltd. (Lot No. MUST-16012417; Sichuan, China) and then dissolved in dimethyl sulfoxide (DMSO). Antibodies against Cyclin B1, Cdc2, BAX, Bcl-2, PARP1, Survivin, PTEN, PI3K, total AKT and GAPDH were all purchased from Proteintech Group, Inc. (Chicago, IL, USA). Antibody against cleaved Caspase3, cleaved Caspase9 and phosphorylated AKT (p-AKT) (ser473) was obtained from Cell Signaling Technology (Danvers, MA, USA). Antibody

Osthole inhibits the proliferation of ESCC cells

The chemical structure of osthole is shown in Fig. 1A. To explore the effect of osthole on ESCC cell proliferation, five ESCC cell lines (KYSE30, KYSE150, KYSE180, KYSE410 and KYSE450) were employed and treated with osthole in different concentrations (0, 20, 40, 80, 120, 160, 200 or 240 μM) for 24 or 48 h. Cell viability was examined using CCK-8 assay for each indicated concentration and time point. As expected, osthole treatment led to reduced viability of ESCC cell in a dose-and

Discussion

Limited clinical approach for treatment remains a major challenge for the patients with ESCC. Increasing evidence suggests that natural agents may represent a new avenue for the development of successful cancer treatments [19]. Such agents have also shown promise in ESCC [20,21]. Osthole, an active compound derived from Fructus Cnidii, has been found widespread application as an anti-inflammatory, anti-allergic, and anti-seizure agent [[11], [12], [13], [14]]. Moreover, both in vitro and in

Acknowledgements

This work was supported by National Natural Science Foundation of China (81402283) and Natural Science Foundation of Liaoning (2015020304).

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