Arachidonic acid suppresses growth of human lung tumor A549 cells through down-regulation of ALDH3A1 expression

doi:10.1016/j.freeradbiomed.2006.01.020

Free Radical Biology and Medicine

Volume 40, Issue 11, 1 June 2006, Pages 1929-1938

https://doi.org/10.1016/j.freeradbiomed.2006.01.020 Get rights and content

Abstract

Expression of aldehyde dehydrogenase 3A1 (ALDH3A1) in certain normal and tumor cells is associated with protection against the growth inhibitory effect of reactive aldehydes generated during membrane lipid peroxidation. We found that human lung tumor (A549) cells, which express high levels of ALDH3A1 protein, were significantly less susceptible to the antiproliferative effects of 4-hydroxynonenal compared to human hepatoma HepG2 or SK-HEP-1 cells that lack ALDH3A1 expression. However, A549 cells became susceptible to lipid peroxidation products when they were treated with arachidonic acid. The growth suppression of A549 cells induced by arachidonic acid was associated with increased levels of lipid peroxidation and with reduced ALDH3A1 enzymatic activity, protein, and mRNA levels. Furthermore, arachidonic acid treatment of the A549 cells resulted in an increased expression of peroxisome proliferator-activated receptor γ (PPARγ), whereas NF-κB binding activity was inhibited. Blocking PPARγ using a selective antagonist, GW9662, prevented the arachidonic acid-mediated reduction of ALDH3A1 expression as well as the growth inhibition of A549 cells, suggesting the central role of PPARγ in these phenomena. The increase in PPARγ and the reduction in ALDH3A1 were also prevented by exposing cells to vitamin E concomitant with arachidonic acid treatment. In conclusion, our data show that the arachidonic acid-induced suppression of A549 cell growth is associated with increased lipid peroxidation and decreased ALDH3A1 expression, which may be due to activation of PPARγ.

Section snippets

Cell cultures

A549 human lung adenocarcinoma cells (ATCC, USA) were seeded in 25-cm² plates at 25,000 cells/cm² and maintained for 24 h in Ham's F-12K medium supplemented with 2 mM glutamine, 1% (v/v) antibiotic/antimycotic solution (medium A), and 10% (v/v) fetal bovine serum (FBS). HepG2 human hepatoma cells (ATCC, USA) were seeded in 25-cm² plates at 20,000 cells/cm² and maintained for 24 h in modified Eagle's medium supplemented with 2 mM glutamine, 1% (v/v) antibiotic/antimycotic solution, 1% (v/v)

Effects of 4-HNE on human lung tumor and hepatoma cells

To investigate the protective effect of ALDH3A1 against lipid peroxidation products in A549, HepG2, and SK-HEP-1 cells, the cells were treated with several different concentrations of 4-HNE and cell growth (determined as number of cells) and viability (determined by flow cytometry) were observed 24 h after the start of 4-HNE treatment. Fig. 1A shows that cell growth in A549 cells was not affected by the 4-HNE treatment. By contrast, both HepG2 and SK-HEP-1 hepatoma cell lines were susceptible

Discussion

In the present study, we report that arachidonic acid suppresses growth in human tumor cell lines. We used human lung tumor cells expressing high levels of ALDH3A1 (A549) and human hepatoma cell lines that lack ALDH3A1 expression (HepG2 and SK-HEP-1) to investigate the role of ALDH3A1 in mediating the inhibitory effects of arachidonic acid on cell proliferation. Our results provide evidence that ALDH3A1 serves as a cell-growth regulator. First, tumor cells expressing ALDH3A1 are resistant to

Acknowledgments

We thank our colleagues for valuable discussions and for critically reading the manuscript. This work was supported by grants from Compagnia di San Paolo, Regione Piemonte, and the University of Turin, Italy, and by EY11490 from the National Institutes of Health, USA.

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Breast cancer chemotherapy can cause side effects due to nonspecific drug delivery, low solubility and fast metabolism of drugs used in conventional therapy. Moreover, the therapeutic effect of the drugs is often reduced by the strengthening of chemoresistance, which occurs via a variety of mechanisms. Different strategies have been developed to reduce multidrug resistance (MDR)-associated gene expressions including the use of surfactants and polymers. In this study superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with conjugated linoleic acid (CLA) reduced the number and viability of cells in comparison with both untreated cells or cells treated with SPIONs alone. This cytostatic effect correlated with the increase of peroxisome proliferator-activated receptors γ (PPARγ). The necrotic cell death induced, as a consequence, an inflammatory process, as evidenced by the decrease of the anti-inflammatory PPARα and increase of pro-inflammatory TNFα and IL-1β. PPARs were examined because CLA is one of their natural ligands. The antitumor effect observed was accompanied by a down-regulation of p-glycoprotein (P-gp), which was the first important discovered efflux transporter belonging to MDR, and of ALDH3A1, an enzyme able to metabolize some drugs, reducing their effects. The down-regulation of P-gp correlated with the increase of cytokines. The ALDH3A1 decrease correlated with the increase of PPARγ. Based on these results, PPARs are molecular mediators of anti-cancer effect of SPIONs functionalized with CLA, being changes in these nuclear receptors correlated with induction of cytotoxicity and inflammation, and decreased ability of cancer cells in blocking anti-cancer drug effect.
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Custard apple (Annona squamosa Linn.) is an edible tropical fruit, and its seeds have been used to treat “malignant sore” (cancer) and other usage as insecticide. A comparison of extraction processes, chemical composition analysis and antitumor activity of A. squamosa seed oil (ASO) were investigated.
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4-Hydroxyhexenal and 4-hydroxynonenal are mediators of the anti-cachectic effect of n-3 and n-6 polyunsaturated fatty acids on human lung cancer cells
2016, Free Radical Biology and Medicine
Citation Excerpt :
The most important finding of the study is that both PUFA families, n-3 and n-6, decreased the proliferation of human lung cancer cells, and were able to prevent the inhibition of muscle cell differentiation induced by lung cancer cells. The observation that AA decreased cancer cell proliferation disagrees with the results of certain other studies [44–46], but confirms previous studies by the present research group evidencing that this n-6 PUFA suppresses the growth of well differentiated human lung tumour cells A549 and of rat hepatocarcinoma cell lines [47–50]. The inhibitory effect of AA on cancer cell proliferation has been inversely related to the ability of cells to metabolize aldehydic products of lipid peroxidation, in particular to the expression of aldehyde dehydrogenase 3A1 (ALDH3A1) [49].
Cachexia, the most severe paraneoplastic syndrome, occurs in about 80% of patients with advanced cancer; it cannot be reverted by conventional, enteral, or parenteral nutrition. For this reason, nutritional interventions must be based on the use of substances possessing, alongside nutritional and energetic properties, the ability to modulate production of the pro-inflammatory factors responsible for the metabolic changes characterising cancer cachexia. In light of their nutritional and anti-inflammatory properties, polyunsaturated fatty acids (PUFAs), and in particular n-3, have been investigated for treating cachexia; however, the results have been contradictory.
Since both n-3 and n-6 PUFAs can affect cell functions in several ways, this research investigated the possibility that the effects of both n-3 and n-6 PUFAs could be mediated by their major aldehydic products of lipid peroxidation, 4-hydroxyhexenal (HHE) and 4-hydroxynonenal (HNE), and by their anti-inflammatory properties. An “in vitro” cancer cachexia model, consisting of human lung cancer cells (A427) and murine myoblasts (C2C12), was used.
The results showed that: 1) both n-3 and n-6 PUFAs reduced the growth of lung cancer cells without causing cell death, increased lipid peroxidation and Peroxisome Proliferator-Activated Receptor (PPAR)α, and decreased TNFα; 2) culture medium conditioned by A427 cells grown in the absence of PUFAs blocked myosin production and the differentiation of C2C12 muscle cells; conversely, muscle cells grown in culture medium conditioned by the same cells in the presence of PUFAs showed myosin expression and formed myotubes; 3) adding HHE or HNE directly to C2C12 cells maintained in culture medium conditioned by A427 cells in the absence of PUFAs stimulated myosin production and myotube formation; 4) putative consensus sequences for (PPARs) have been found in genes encoding fast isoforms of myosin heavy chain, by a bioinformatics approach.
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Citation Excerpt :
In support of this concept, it has been recently shown that knock-in of ALDH2∗2 in mice heighten nociception, which could be attenuated upon treatment with Alda-1, suggesting that treatment with Alda-1 could increase pain tolerance in individuals carrying the ALDH2 point mutation [140]. The subfamily of ALDH3 enzymes such as ALDH3A1 are encoded by the chromosome 17p11.2 [141] and have been reported to have multiple functions including the maintenance of hematopoietic stem cells, protection of the eye from UV radiation [142], regulating cell proliferation and lipid-peroxidation mediated growth inhibition [143], in attenuating ROS induced protein modification [144], preventing DNA damage and apoptosis [145] and providing resistance against chemotherapeutic drugs [143,146–152]. Since ALDH3A1 is expressed abundantly in corneal epithelium [153] it is considered to play a role in maintaining the corneal function of light transparency and refraction [154].
Extensive research has shown that increased production of reactive oxygen species (ROS) results in tissue injury under a variety of pathological conditions and chronic degenerative diseases. While ROS are highly reactive and can incite significant injury, polyunsaturated lipids in membranes and lipoproteins are their main targets. ROS-triggered lipid-peroxidation reactions generate a range of reactive carbonyl species (RCS), and these RCS spread and amplify ROS-related injury. Several RCS generated in oxidizing lipids, such as 4-hydroxy trans-2-nonenal (HNE), 4-oxo-2-(E)-nonenal (ONE), acrolein, malondialdehyde (MDA) and phospholipid aldehydes have been shown to be produced under conditions of oxidative stress and contribute to tissue injury and dysfunction by depleting glutathione and other reductants leading to the modification of proteins, lipids, and DNA. To prevent tissue injury, these RCS are metabolized by several oxidoreductases, including members of the aldo–keto reductase (AKR) superfamily, aldehyde dehydrogenases (ALDHs), and alcohol dehydrogenases (ADHs). Metabolism via these enzymes results in RCS inactivation and detoxification, although under some conditions, it can also lead to the generation of signaling molecules that trigger adaptive responses. Metabolic transformation and detoxification of RCS by oxidoreductases prevent indiscriminate ROS toxicity, while at the same time, preserving ROS signaling. A better understanding of RCS metabolism by oxidoreductases could lead to the development of novel therapeutic interventions to decrease oxidative injury in several disease states and to enhance resistance to ROS-induced toxicity.

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Original ContributionArachidonic acid suppresses growth of human lung tumor A549 cells through down-regulation of ALDH3A1 expression

Abstract

Section snippets

Cell cultures

Effects of 4-HNE on human lung tumor and hepatoma cells

Discussion

Acknowledgments

Arch. Biochem. Biophys.

Biochem. Pharmacol.

Biochem. Pharmacol.

J. Biol. Chem.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

J. Biol. Chem.

Biochem. Pharmacol.

Chem. Biol. Interact.

Chem. Biol. Interact.

Toxicology

Free Radic. Biol. Med.

Exp. Cell Res.

Free Radic. Biol. Med.

Biochimie

J. Biol. Chem.

Gastroenterology

Cell Signalling

Hepatology

Cancer Cell

J. Surg. Res.

Atherosclerosis

Neoplasia

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

J. Biol. Chem.

Original Contribution
Arachidonic acid suppresses growth of human lung tumor A549 cells through down-regulation of ALDH3A1 expression