T63, a new 4-arylidene curcumin analogue, induces cell cycle arrest and apoptosis through activation of the reactive oxygen species–FOXO3a pathway in lung cancer cells

doi:10.1016/j.freeradbiomed.2012.10.537

Free Radical Biology and Medicine

Volume 53, Issue 12, 15 December 2012, Pages 2204-2217

https://doi.org/10.1016/j.freeradbiomed.2012.10.537 Get rights and content

Abstract

Curcumin (diferuloylmethane) is a natural polyphenol product of the plant Curcuma longa and has a diversity of antitumor activities. T63, a new 4-arylidene curcumin analogue, was reported to inhibit proliferation of lung cancer cells. However, its precise molecular antitumor mechanisms have not been well elucidated. Here, we showed that T63 could significantly inhibit the proliferation of A549 and H460 human lung cell lines via induction of G0/G1 cell cycle arrest and apoptosis. We found that the reactive oxygen species (ROS)-activated FOXO3a cascade plays a central role in T63-induced cell proliferation inhibition. Mechanistically, enhancement of ROS production by T63 induced FOXO3a expression and nuclear translocation through activation of p38MAPK and inhibition of AKT, subsequently elevating the expression of FOXO3a target genes, including p21, p27, and Bim, and then increased the levels of activated caspase-3 and decreased the levels of cyclin D1. Moreover, the antioxidant N-acetylcysteine markedly blocked the above effects, and small interfering RNA-mediated knockdown of FOXO3a also significantly decreased T63-induced cell cycle arrest and apoptosis. In vivo experiments showed that T63 significantly suppressed the growth of A549 lung cancer xenograft tumors, associated with proliferation suppression and apoptosis induction in tumor tissues, without inducing any notable major organ-related toxicity. These data indicated that the novel curcumin analogue T63 is a potent antitumor agent that induces cell cycle arrest and apoptosis and has significant therapeutic potential for lung cancer.

Highlights

► T63, a curcumin analogue, induced G0/G1 arrest and apoptosis in lung cancer cells. ► T63 treatment rapidly increased reactive oxygen species levels. ► Activation of FOXO3a is essential for T63-induced cell cycle arrest and apoptosis. ► T63 significantly suppressed the growth of A549 lung cancer xenograft tumors. ► T63 treatment showed no notable organ-related toxicity in vivo.

Section snippets

Materials

T63 and curcumin were synthesized at our laboratory, dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mM stock solution, and stored at −20 °C. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide, 3,3′-diaminobenzidine, SB203580, LY294002, diphenylene iodonium (DPI), rotenone, allopurinol, PEG–catalase (PEG-CAT), PEG–superoxide dismutase (PEG-SOD), and N-acetylcysteine (NAC) were obtained from Sigma (Poole, UK). 4,6-Diamidino-2-phenylindole (DAPI), the

Effects of T63 on cell viability

We first determined the effects of T63 (Fig. 1A) on cell proliferation of lung cancer cells. A549 and H460 cells were treated with T63 or curcumin (0.01 to 10 μM) for 24 h, and then the viability was determined by MTT assay. As shown in Fig. 1B, T63 significantly suppresses proliferation of A549 and H460 cells within a 24-h treatment period. More important, the inhibitory effects were observed at a dose of 1 μM, whereas curcumin showed no significant inhibitory effect upon A549 or H460 cell

Discussion

The ability of cancer cells to avoid apoptosis has been identified as one of the major mechanisms for development of cancers [36]. Induction of cell cycle arrest and cell apoptosis in cancer cells is a critical feature of chemotherapeutic agents. Curcumin has been shown to inhibit proliferation of various human tumor cells, including lung, colon, breast, myeloma, leukemia, and prostate carcinoma [21], [37]. The chemotherapeutic action of curcumin might be due to its ability to induce apoptosis

Acknowledgments

This work was supported by grant from the MOST of China (863 Program 2008AA02Z304), the National Natural Science Foundation of China (Grants 30973619, 31101071, 81071712, 30873032, and 81172931), and the National Basic Research Program of China (973 Program 2011CB9358003).

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Original ContributionT63, a new 4-arylidene curcumin analogue, induces cell cycle arrest and apoptosis through activation of the reactive oxygen species–FOXO3a pathway in lung cancer cells

Abstract

Highlights

Section snippets

Materials

Effects of T63 on cell viability

Discussion

Acknowledgments

Lung Cancer

J. Biol. Chem.

J. Biol. Chem.

Cell

Cell

Cell

Cell

Life Sci.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Biochem. Pharmacol

Cell

Cancer Lett.

Cancer Cell

Cancer Cell

Free Radic. Biol. Med.

Biochim. Biophys. Acta

Free Radic. Biol. Med.

Free Radic. Biol. Med

Mol. Cell. Endocrinol.

Free Radic. Biol. Med.

J. Biol. Chem.

J. Biol. Chem.

Biochimie

Free Radic. Biol. Med.

Free Radic. Biol. Med

Free Radic. Biol. Med.

Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths

CA Cancer J. Clin

Mechanisms of resistance to therapy and tumor cell survival

Curr. Opin. Hematol.

Tumor intracellular redox status and drug resistance—serendipity or a causal relationship?

Curr. Pharm. Des

Sense and sensitivity: FOXO and ROS in cancer development and treatment

Antioxid. Redox Signaling

Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach?

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FOXO3a mediates the androgen-dependent regulation of FLIP and contributes to TRAIL-induced apoptosis of LNCaP cells

Oncogene

Original Contribution
T63, a new 4-arylidene curcumin analogue, induces cell cycle arrest and apoptosis through activation of the reactive oxygen species–FOXO3a pathway in lung cancer cells