Erianin inhibits indoleamine 2, 3-dioxygenase –induced tumor angiogenesis

doi:10.1016/j.biopha.2017.01.090

Biomedicine & Pharmacotherapy

Volume 88, April 2017, Pages 521-528

https://doi.org/10.1016/j.biopha.2017.01.090 Get rights and content

Abstract

Tumor angiogenesis is the key process in tumor growth and metastasis, and transfers essential nutrients for solid tumor. Inhibition of tumor angiogenesis has been recognized as a more effective anti-cancer strategy for NSCLC and has acquired certain therapeutic effects. IDO has non-immune functions including regulating tumor angiogenesis and IDO dysregulation in cancer pathogenesis has been valued. Erianin is a natural product isolated from Dendrobium chrysotoxum Lindl. The antitumor activity of erianin in many kinds of cancers had been demonstrated in previous studies.

In this study, we demonstrated that IDO could promote the attachment of 2LL cells, the ability of migration, invasion and VM formation, as well as the tubules forming ability of HUVECs. We also find that erianin suppressed expression and enzyme ability of IDO and erianin could inhibit IDO-induced metastasis and invasion ability of 2LL cells significantly. Erianin not only blocked IDO-induced tube formation of HUVECs, but also suppressed VM formation of 2LL-IDO cells. What’s more, we examined that Erianin might play its role in angiogenesis through down-regulating phosphorylation of JAK2/STAT3, inhibiting its downstream target genes MMP-2/-9 and some inflammatory mediators (COX-2, HIF-1α and IL-6), which were all induced by IDO.

All these results indicated that erianin had anti-angiogenesis ability and could inhibit the expresison of IDO to prevent and treat the malignant tumors.

Introduction

There are few obvious early symptoms of lung cancer, so the patients are diagnosed at middle and late stage, leading to the poor prognosis with low quality of life [1]. With platinum-based chemotherapy is the standard first-line treatment of advanced non-small cell lung cancer. Angiogenesis is a very important biological processes. Many studies found that angiogenesis has a close relationship with non-small cell lung cancer development, metastasis and prognosis [2].

Erianin is natural products of low molecular extracted from the Dendrobium Chrysostom [3]. In traditional Chinese medicine, Erianin is often used as antipyretic and analgesic agent. Erianin belongs to bibenzyl derivatives in structure, with the activity of anti-virus [3], anti-bacterial [4], as well as anti-benign prostatic hyperplasia [5].

Angiogenesis is the process of formation of new blood vessels, is a sign of tumor processes. Angiogenesis plays an important role in many physiological processes, including embryonic development, tissue repair of postoperative and Posttraumatic. It can also be used as a marker in wound healing, cancer, ischemic and inflammatory diseases [6], [7], [8].

Indole amine 2,3-dioxygenase(IDO)is the rate-limiting enzyme in the pathway of tryptophan (TRP) along the dog urine ammonia acid (KYN) in the metabolic process. Almost 99% TRP metabolizes through dog urine acid pathway [9]. Li, etc. [10] co-cultured fibroblast cells of overexpression of IDO and vascular endothelial cells in vitro, and found micro-vascular endothelial cells, and untreated fibroblasts do not encourage micro-vascular endothelial cells, indicating that IDO can directly cause the endothelial cells of the blood vessel formation. Dog Urine ammonia acid was added to reverse the IDO-mediated angiogenesis, suggesting that tryptophan depletion may be one of the mechanisms of stimulating angiogenesis [11]. The overexpression of IDO in ovarian cancer cells can result in a significant increase in the number of new blood vessels, confirmed that immune tolerance is not the only function of IDO, IDO is also able to induce tumor angiogenesis [12], playing a non-immunological role, providing new targets for therapeutic angiogenesis.

Erianin can has the anti-angiogenic effect by inducing apoptosis of vascular endothelial cells [13]. IDO is involved in various stages of cancer development, and the current research on IDO focuses on its immunological function. Recently, it is found that the immune regulation function is not the only function of IDO, and IDO plays an important role in the invasion, metastasis and angiogenesis.

So we use the application of in vitro experiment methods to study whether Erianin can inhibit lung cancer cell invasion, metastasis and angiogenesis by the regulation of IDO expression, and provide a theoretical basis for lung cancer anti-angiogenesis therapy.

Section snippets

Reagents

The powder of Erianin was dissolved in dimethyl sulfoxide (DMSO), with a concentration of 1 mM solution. 1-Methyl-d-tryptophan (1-d-MT)(Sigma-Aldrich)was prepared into 200 mM solution under alkaline conditions, and then adjusted pH to 7.4 by HCl. They were all stored at −20 °C.The blue MTT formazan precipitate was dissolved in 100 μl of DMSO.

Cell culture

The mouse Lewis lung cancer cell lines(2LL)was generously donated by Longhua hospital (Shanghai, China) and 2LL-IDO cells which express human IDO gene were

Statistical analysis

Statistical analyses were performed using GraphPad Prism 5.0 software (GraphPad Software Inc, USA)with the two-tailed Student’s t-test. Significance was determined at the 95% confidence interval. All data were expressed as the mean ± standard deviation (SD) from a representative experiment.

Verification of overexpression of IDO in lewis lung cancer cells in mice

Firstly, in order to use the 2LL-IDO cells in the following experiments, we verified the expression of IDO gene in 2LL-IDO cells. Because 2LL-EGFP-IDO cells with green fluorescent protein, the fluorescence of the cells was observed under fluorescence microscope. As shown in Fig. 1A, 2LL-EGFP-IDO cells can emit green fluorescence under blue excitation light. Next, we used Western blot and qPCR to detect the expression of IDO in 2LL-IDO cells at the protein and mRNA levels, respectively. The

Discussion

The study of tumor angiogenesis is of great value in the early diagnosis, treatment and prognosis of tumor [16]. Dendrobium has high medicinal value, and its efficacy has been the focus of attention over the world [17]. As one of the effective components of Dendrobium, Erianin has great anti-tumor and anti-angiogenesis effect in vitro and in vivo [3]. We studied the effect of IDO on the angiogenesis of Lewis lung cancer in mice with IDO as the breakthrough point, and expounds the Erianin can

Conflicts of interest

The authors have declared that no competing interests exist.

Funding

The authors have no support or funding to report.

References (19)

T. Ng et al.
Antioxidative activity of natural products from plants
Life Sci.
(2000)
O. Takikawa
Tryptophan degradation in mice initiated by indoleamine 2: 3-dioxygenase
J. Biol. Chem.
(1986)
Y. Li
Local expression of indoleamine 2: 3-dioxygenase protects engraftment of xenogeneic skin substitute
J. Invest. Dermatol.
(2006)
Y.-Q. Gong
In vivo and in vitro evaluation of erianin: a novel anti-angiogenic agent
Eur. J. Cancer
(2004)
Group et al.
Chemotherapy in addition to supportive care improves survival in advanced non?Small-Cell lung cancer: a systematic review and meta-Analysis of individual patient data from 16 randomized controlled trials
J. Clin. Oncol.
(2008)
A. Yuan
Tumor angiogenesis correlates with histologic type and metastasis in non-small-cell lung cancer
Am. J. Respir. Crit. Care Med.
(1995)
E. Kámory et al.
Isolation and antibacterial activity of marchantin A: a cyclic bis (bibenzyl) constituent of Hungarian Marchantia polymorpha
Planta Med.
(1995)
J. Dekanski
Anti-prostatic activity of bifluranol, a fluorinated bibenzyl
Br. J. Pharmacol.
(1980)
J. Folkman
Angiogenesis in cancer: vascular, rheumatoid and other disease
Nat. Med.
(1995)

There are more references available in the full text version of this article.

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¹: These authors contributed equally to this article and should be considered as co-first authors.

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Erianin inhibits indoleamine 2, 3-dioxygenase –induced tumor angiogenesis

Abstract

Introduction

Section snippets

Reagents

Cell culture

Statistical analysis

Verification of overexpression of IDO in lewis lung cancer cells in mice

Discussion

Conflicts of interest

Funding

Life Sci.

J. Biol. Chem.

J. Invest. Dermatol.

Eur. J. Cancer

Chemotherapy in addition to supportive care improves survival in advanced non?Small-Cell lung cancer: a systematic review and meta-Analysis of individual patient data from 16 randomized controlled trials

J. Clin. Oncol.

Tumor angiogenesis correlates with histologic type and metastasis in non-small-cell lung cancer

Am. J. Respir. Crit. Care Med.

Isolation and antibacterial activity of marchantin A: a cyclic bis (bibenzyl) constituent of Hungarian Marchantia polymorpha

Planta Med.

Anti-prostatic activity of bifluranol, a fluorinated bibenzyl

Br. J. Pharmacol.

Angiogenesis in cancer: vascular, rheumatoid and other disease

Nat. Med.