Polygonatum cyrtonema lectin induces apoptosis and autophagy in human melanoma A375 cells through a mitochondria-mediated ROS–p38–p53 pathway

doi:10.1016/j.canlet.2008.09.042

Cancer Letters

Volume 275, Issue 1, 8 March 2009, Pages 54-60

https://doi.org/10.1016/j.canlet.2008.09.042 Get rights and content

Abstract

Polygonatum cyrtonema lectin (PCL), a mannose-binding lectin, has been reported to induce cytotoxicity and apoptosis. Herein, we demonstrated that PCL-induced apoptosis and autophagy in A375 cells. The apoptotic mechanism was that PCL treatment regulated Bax, Bcl-xL and Bcl-2 proteins, leading to mitochondrial depolarization, cytochrome c release and caspase activation. Subsequently, we found that PCL treatment abrogated glutathione antioxidant system and induced mitochondria to generate ROS accumulation, resulting in p38–p53 activation. Moreover, we confirmed that the ROS–p38–p53 pathway was involved in PCL-induced autophagy. In conclusion, these results indicate that PCL induces apoptosis and autophagy via a mitochondrial-mediated ROS–p38–p53 pathway.

Introduction

Plant lectins, a class of carbohydrate-binding non-immune origin proteins, are known to possess the ability to agglutinate cells or precipitate polysaccharides and glycoconjugates [1]. In the past few years, they have been used to differentiate malignant tumors from benign and the degree of glycosylation associated with metastasis [2]. In addition, several plant lectins, especially mistletoe lectins have been adopted for alternative cancer therapy for several years [3].

Of note, cancer is associated with the programmed cell death (PCD), which plays a crucial role in the development and homeostasis of multicellular organisms [4]. Apoptosis, type I PCD, is characterized by condensation of the cytoplasm and nucleus, DNA fragmentation, chromatin merging in the nuclear periphery, cell contraction, dynamic membrane blebbing, and cell phagocytosis [5]. Autophagy, type II PCD, is independent of phagocytes and differs from apoptosis by the presence of autophagosomes, autolysosomes, and an intact nucleus in the cell [6]. Autophagy is not only a survival response to either growth factor or nutrient deprivation but an important mechanism for tumor cell suicide [7]. More importantly, some investigations have demonstrated that the co-regulation of both apoptosis and autophagy can participate in mammalian cell death [8]. Other reports have further pointed out that apoptosis and autophagy may be interconnected and even simultaneously regulated by the same trigger [9].

Previous studies have reported that several plant lectins can be involved in apoptosis and autophagy in tumor cells [10], [11], [12]. However, the apoptotic and/or autophagic mechanisms of them remain uncertain. PCL, a mannose/sialic acid-binding lectin, was reported for its inhibitory and apoptosis-inducing effects on the growth of cancer cells [13]. The precise mechanism by which PCL induces tumor cell death is however still only rudimentarily understood. In this study, we report for the first time that PCL can simultaneously induce apoptosis and autophagy in tumor cells. Moreover, we further explore that the molecular mechanism of apoptosis and autophagy induced by PCL is a mitochondria-mediated ROS–p38–p53 pathway.

Section snippets

Cell culture and reagents

The normal human melanocytes and melanoma A375 cells were both obtained from Medical Sciences Center of West China in Sichuan University. They were routinely cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100 U/ml streptomycin, 100 U/ml penicillin, and 2 mm l-glutamine in a humidified cell incubator with an atmosphere of 5% CO₂ at 37 °C.

Polygonatum cyrtonema lectin (PCL) was purified as previously described [14]. Fetal bovine serum (FBS),

PCL induces apoptosis and autophagy in A375 cells

PCL caused remarkable inhibition of A375 cell growth in a time- and dose-dependent manner, and treatment with 15 μg/ml PCL for 24 h resulted in almost 50% inhibition (Fig. 1A, left). In contrast, only a small percentage of cell death was found in the normal melanocytes after treatment with 15 μg/ml PCL for 24 h (Fig. 1A, right). The apoptotic ultrastructural alterations were observed under electron microscope. Compared to control cells, PCL-treated A375 cells revealed nuclear fragmentation (Fig. 1

Discussion

In the current study, we demonstrated that PCL was remarkable inhibitory toward A375 cells (IC₅₀ = 15 μg/ml, 24 h), but was low toxic to the normal melanocytes, suggesting PCL bears a greater susceptibility of the malignant cells. Thus, PCL would be utilized as a potential candidate agent for its susceptibility of cancer cells.

In addition, we demonstrated that PCL induced both apoptosis and autophagy in A375 cells. Previous studies have pointed out several plant lectins can induce apoptosis in

Conflicts of interest

The authors declare that they have no competing interests.

Acknowledgements

This work was supported in part by grants from the National Natural Science Foundation of China (General Programs: no. 30270331 and no. 30670469), and Director Fund of State Key Laboratory of Oral Diseases (Sichuan University).

References (28)

N. Sharon
Carbohydrate-specific reagents and biological recognition molecules
J. Biol. Chem.
(2007)
B. Levine et al.
Development by self-digestion: molecular mechanisms and biological functions of autophagy
Dev. Cell
(2004)
L. Xue et al.
Autophagy is activated by apoptotic signalling in sympathetic neurons: an alternative mechanism of death execution
Mol. Cell Neurosci.
(1999)
Z. Liu et al.
A mannose-binding lectin from Sophora flavescens induces apoptosis in HeLa cells
Phytomedicine
(2008)
G. Seifert et al.
Molecular mechanisms of mistletoe plant extract-induced apoptosis in acute lymphoblastic leukemia in vivo and in vitro
Cancer Lett.
(2008)
T. Mosmann
Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
J. Immunol. Methods
(1983)
N.E. Saris et al.
Interference of calmidazolium with measurement of mitochondrial membrane potential using the tetraphenylphosphonium electrode or the fluorescent probe rhodamine 123
Anal. Biochem.
(2004)
Y. Cheng et al.
Apoptosis-suppressing and autophagy-promoting effects of calpain on oridonin-induced A375 cell death
Arch. Biochem. Biophys.
(2008)
M.S. Kim et al.
Involvement of hydrogen peroxide in mistletoe lectin-II-induced apoptosis of myeloleukemic U937 cells
Life Sci.
(2003)
R.C. Scott et al.
Direct induction of autophagy by Atg1 inhibits cell growth and induces apoptotic cell death
Curr. Biol.
(2007)

D. Kessel et al.

Initiation of apoptosis and autophagy by the Bcl-2 antagonist HA14-1

Cancer Lett.

(2007)

E. Gorelik et al.

On the role of cell surface carbohydrates and their binding proteins (lectins) in tumor metastasis

Cancer Metast. Rev.

(2001)

K. Schumacher et al.

Influence of postoperative complementary treatment with lectin-standardized mistletoe extract on breast cancer patients: a controlled epidemiological multicentric retrospective cohort study

Anticancer Res.

(2003)

Y. Kondo et al.

The role of autophagy in cancer development and response to therapy

Nat. Rev. Cancer

(2005)

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Polygonatum cyrtonema lectin induces apoptosis and autophagy in human melanoma A375 cells through a mitochondria-mediated ROS–p38–p53 pathway

Abstract

Introduction

Section snippets

Cell culture and reagents

PCL induces apoptosis and autophagy in A375 cells

Discussion

Conflicts of interest

Acknowledgements

J. Biol. Chem.

Dev. Cell

Mol. Cell Neurosci.

Phytomedicine

Cancer Lett.

J. Immunol. Methods

Anal. Biochem.

Arch. Biochem. Biophys.

Life Sci.

Curr. Biol.

Cancer Lett.

On the role of cell surface carbohydrates and their binding proteins (lectins) in tumor metastasis

Cancer Metast. Rev.

Influence of postoperative complementary treatment with lectin-standardized mistletoe extract on breast cancer patients: a controlled epidemiological multicentric retrospective cohort study

Anticancer Res.

The role of autophagy in cancer development and response to therapy

Nat. Rev. Cancer