Elsevier

European Journal of Pharmacology

Volume 795, 15 January 2017, Pages 150-159
European Journal of Pharmacology

Molecular and cellular pharmacology
Propofol attenuates pancreatic cancer malignant potential via inhibition of NMDA receptor

https://doi.org/10.1016/j.ejphar.2016.12.017Get rights and content

Abstract

Propofol is a commonly used intravenous anesthetic, and could attenuate cancer cells malignant potential via inhibiting hypoxia-inducible factor-1α (HIF-1α) expression. However, the mechanism is still inclusive. In the present study, we mainly focus on the mechanism by which propofol down-regulated HIF-1α expression and malignant potential in pancreatic cancer cells. Human pancreatic cancer cells (Miapaca-2 and Panc-1) in vitro and murine pancreatic cancer cell (Panc02) in vivo were used to assess the effect of propofol on vascular endothelial growth factor (VEGF) expression and migration of pancreatic cancer cells. Propofol inhibited cells migration, expression of VEGF and HIF-1α, phosphorylation of extracellular regulated protein kinases (ERK), AKT, Ca2+/calmodulin dependent protein kinases II (CaMK II), and Ca2+ concentration in a concentration-dependent manner (5, 25, 50, 100 μM). Furthermore, MK801, an inhibitor of NMDA receptor, and KN93, an inhibitor of CaMK II, could inhibit the expression of VEGF, HIF-1a, p-AKT, p-ERK, p-CaMK II in vitro, growth of tumor and VEGF expression in vivo, which were similar to the effect of propofol. In addition, the anti-tumor effect of propofol could be counteracted by rapastinel, an activator of NMDA receptor. Our study indicated that propofol suppressed VEGF expression and migration ability of pancreatic cancer cells in vitro and in vivo, probably via inhibiting NMDA receptor.

Introduction

Cancer remains one of the main causes of death worldwide and surgery is the prior option for most solid tumor treatment. However surgical stress could affect the immune and neuroendocrine systems, and induce inadvertent seeding of tumor cells during surgery (Boomsma et al., 2010, Camara et al., 2006, Mori et al., 1996, Tsuchiya et al., 2003), thus affecting the prognosis of the patients. Despite great progress made in surgical skills and perioperative care, the overall survival rate in patients with pancreatic cancer is still low. Moreover, pancreatic cancer surgery duration is usually as long as 6–7 h (Gangavatiker et al., 2011, Okano et al., 2015), which allows the anesthesiologist to possess more time to participate in the treatment procedure intra-operatively.

Elevation of hypoxia-inducible factor 1α (HIF-1α) expression has been found in a variety of human cancers (Talks et al., 2000). As a directly master control of the expression of numbers of genes, HIF-1α up-regulation is related to tumor growth (Terraneo et al., 2010, Wang et al., 2011), vascularization (Ban et al., 2010) and migration (Burrows et al., 2011), thus leading to poor patient outcome. Moreover, studies have indicated that ERK (Liu et al., 2010) and AKT (Huang et al., 2014) lies upstream of HIF-1α, and activation of ERK or AKT could activate HIF-1α (Huang et al., 2014, Liu et al., 2010).

Large amount of clinical research indicate that anesthesia methods could influence prognosis of cancer patients (Biki et al., 2008, Christopherson et al., 2008, Exadaktylos et al., 2006, Lin et al., 2011). However, (1) all this data are retrospective and therefore provide very weak evidence, and (2) there are also a large number of studies that cannot show any effect of anesthetics on cancer. Recent studies indicated that anesthetics could affect cancer cells malignant potential (Benzonana et al., 2013, Huang et al., 2014). Inhalation anesthetics and propofol were reported to play an opposite role on cancer biology. Isoflurane augmented tumor growth in human cancer cells, while propofol inhibited cancer cells proliferation (Benzonana et al., 2013; Huang et al., 2014). Propofol is a commonly used intravenous anesthetic in clinic. It has been proposed that propofol could inhibit cancer cells malignant potential via inhibiting HIF-1α expression (Huang et al., 2014). However, the mechanism has not been elucidated. In the present study, we mainly focus on the mechanism by which propofol down-regulates HIF-1α expression and malignant potential in pancreatic cancer cells.

Section snippets

Cell culture and reagent

Miapaca-2, Panc-1 and Panc02, purchased from the American Type Culture Collection (ATCC), were cultured in DMEM with 10% fetal bovine serum at 37 °C in a humidified 5% CO2 incubator. Cells were sub-cultured when reaching 90% confluence.

KN93, an inhibitor of Ca2+/calmodulin dependent protein kinases (CaMK II), MK801, an inhibitor of N-Methyl-D-aspartate (NMDA) receptor, and rapastinel, an activator of NMDA receptor, were purchased for Sigma (St. Louis, MO). The substances employed in the present

Propofol inhibits VEGF expression and cells migration in Miapaca-2 and Panc-1 cells

In Miapaca-2 and Panc-1 cells, propofol inhibited VEGF expression (Fig. 1A, B) and migration (Fig. 1C) in a concentration-dependent manner. Compared with control group, incubation of cells with 50 μM propofol significantly attenuated VEGF expression and cells migration in Miapaca-2 and Panc-1 cells (Fig. 1). Moreover, the propofol solvent dimethyl sulfoxide (DMSO) did not affect VEGF expression and cells migration.

Propofol inhibits HIF-1α expression, ERK and AKT phosphorylation in Miapaca-2 and Panc-1 cells

In Miapaca-2 and Panc-1 cells, propofol inhibited HIF-1α expression (Fig. 2A, B),

Discussion

In the present study we proposed that propofol via inhibiting NMDA receptor, attenuated intracellular Ca2+ concentration, CaMK II, ERK, AKT phosphorylation and HIF-1α expression, thus suppressing VEGF expression, cells migration and the growth of tumor.

HIF-1α has been indicated to activate a spectrum of downstream genes to promote cell proliferation (Terraneo et al., 2010, Wang et al., 2011), angiogenesis (Ban et al., 2010) and migration (Burrows et al., 2011). Upregulation of HIF-1α expression

Conclusions

In summary, the present study indicated that propofol attenuated Ca2+ concentration, inhibited CaMK II, ERK and AKT phosphorylation, down-regulated HIF-1α expression, and eventually inhibited VEGF expression, cells migration and the growth of tumor. The anti-tumor effect of propofol may be achieved via inhibition of NMDA receptor.

Declaration

Xiangyuan Chen, Qichao Wu, Li You, Sisi Chen, Minmin Zhu and Changhong Miao have none conflicts of interest.

Acknowledgements

We thank Zhen Wang (Fudan University Shanghai Cancer Center, Shanghai, China) for assistance in experimental technologies.

This project is supported by Shanghai Charity Cancer Research Center (No. HYXH1407).

References (41)

  • K.L. Talks et al.

    The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages

    Am. J. Pathol.

    (2000)
  • Y. Tsuchiya et al.

    Increased surgical stress promotes tumor metastasis

    Surgery

    (2003)
  • C. Wang et al.

    A novel endogenous human CaMKII inhibitory protein suppresses tumor growth by inducing cell cycle arrest via p27 stabilization

    J. Biol. Chem.

    (2008)
  • D. Zhang et al.

    GABAergic signaling facilitates breast cancer metastasis by promoting ERK1/2-dependent phosphorylation

    Cancer Lett.

    (2014)
  • B.A. Orser et al.

    Inhibition by propofol (2,6 di-isopropylphenol) of the N-methyl-D-aspartate subtype of glutamate receptor in cultured hippocampal neurones

    Br. J. Pharm.

    (1995)
  • L.L. Benzonana et al.

    Isoflurane, a commonly used volatile anesthetic, enhances renal cancer growth and malignant potential via the hypoxia-inducible factor cellular signaling pathway in vitro

    Anesthesiology

    (2013)
  • B. Biki et al.

    Anesthetic technique for radical prostatectomy surgery affects cancer recurrence: a retrospective analysis

    Anesthesiology

    (2008)
  • M.F. Boomsma et al.

    Breast cancer surgery-induced immunomodulation

    J. Surg. Oncol.

    (2010)
  • A. Britschgi et al.

    Calcium-activated chloride channel ANO1 promotes breast cancer progression by activating EGFR and CAMK signaling

    Proc. Natl. Acad. Sci. USA

    (2013)
  • N. Burrows et al.

    GDC-0941 inhibits metastatic characteristics of thyroid carcinomas by targeting both the phosphoinositide-3 kinase (PI3K) and hypoxia-inducible factor-1alpha (HIF-1alpha) pathways

    J. Clin. Endocrinol. Metab.

    (2011)
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