ISSN 1214-0287 (on-line), ISSN 1214-021X (printed)
J Appl Biomed
Volume 11 (2013), No 2, p 93-103
DOI 10.2478/v10136-012-0033-8

Alpha-tomatine activates cell cycle checkpoints in the absence of DNA damage in human leukemic MOLT-4 cells

Jana Kudelova, Martina Seifrtova, Lenka Sucha, Pavel Tomsik, Radim Havelek, Martina Rezacova

Address: Martina Rezacova, Department of Med. Biochemistry, Medical Faculty in Hradec Kralove, Simkova 870, 500 38 Hradec Kralove 1, Czech Republic
rezacovam@lfhk.cuni.cz

Received 18th October 2012.
Revised 20th December 2012.
Published online 8th January 2013.

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SUMMARY
Alpha-tomatine is a major glycoalkaloid found in the roots, leaves, stems and fruit of tomatoes Lycopersicon esculentum. Recently, alpha-tomatine has been recognized as a potential anticancer drug. In the present study, we identified the signaling cascades involved in the antitumor effect of alpha-tomatine on MOLT-4 leukemic cells. Alpha-tomatine inhibited the proliferation and decreased the viability of MOLT-4 cells in a dose-dependent manner. An increase in the activity of caspases 9 and 3/7 was not observed. However, an increase in the amount of p53 and its phosphorylation on serine 15, as well as an increased amount of mitochondrial protein PUMA was detected 4 and 24 h after exposure to alpha-tomatine at a concentration of 1–3 micromol/l. Inhibition of the proliferation of MOLT-4 cells by alpha-tomatine is also associated with an increase in p21WAF1/CIP1 and the activation of Chk2. The comet assay did not detect significant amounts of single or double DNA strand breaks in cells treated with alpha-tomatine at concentrations of 0.1-9 mmol/l. Our results thus contribute to the understanding of the anticancer action of alpha-tomatine.

KEY WORDS
alpha-tomatine; DNA damage; p53; PUMA; leukemia

REFERENCES
Bai L, Zhu WG. P53: Structure, unction and therapeutic applications. J. Cancer Mol. 2: 141-153, 2006.

Bartek J, Lukas J. Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell. 3: 421-429, 2003.
[CrossRef]

Berger J. The age of biomedicine: current trends in traditional subjects. J Appl Biomed 9: 57-61, 2011.
[CrossRef] [JAB]

Bergers WW, Alink GM. Toxic effect of the glycoalkaloids solanine and tomatine on cultured neonatal rat heart cells. Toxicol Lett. 6: 29-32, 1980.
[CrossRef]

Bogner C, Leber B, Andrews DW. Apoptosis: embedded in membranes. Curr Opin Cell Biol. 22: 845-851, 2010.
[CrossRef] [PubMed]

Calini V, Urani C, Camatini M. Comet assay evaluation of DNA single- and double-strand breaks induction and repair in C3H10T1/2 cells. Cell Biol Toxicol. 18: 369-379, 2002.
[CrossRef] [PubMed]

Cazzalini O, Scovassi AI, Savio M, Stivala LA, Prosperi E. Multiple roles of the cell cycle inhibitor p21(CDKN1A) in the DNA damage response. Mutat Res. 704: 12–20, 2010.
[CrossRef] [PubMed]

Chao MW, Chen CH, Chang YL, Teng CM, Pan SL. Alpha-Tomatine-Mediated Anti-Cancer Activity In Vitro and In Vivo through Cell Cycle- and Caspase-Independent Pathways. PLoS One. 7: e44093, 2012.
[CrossRef] [PubMed]

Chiu FL, Lin JK. Tomatidine inhibits iNOS and COX-2 through suppression of NF-kappaB and JNK pathways in LPS-stimulated mouse macrophages. FEBS Lett. 582: 2407-2412, 2008.
[CrossRef] [PubMed]

Choi SH, Ahn JB, Kozukue N, Kim HJ, Nishitani Y, Zhang L, Mizuno M, Levin CE, Friedman M. Structure-activity relationships of alpha-, beta(1)-, gamma-, and delta-tomatine and tomatidine against human breast (MDA-MB-231), gastric (KATO-III), and prostate (PC3) cancer cells. J Agric Food Chem. 60: 3891– 3899, 2012.
[CrossRef]

Collins AR, Dusinska M, Gedik CM, Stetina R. Oxidative damage to DNA: do we have a reliable biomarker? Environ Health Perspect. 104(Suppl 3): 465-469, 1996.
[PubMed]

Erlacher M, Michalak EM, Kelly PN, Labi V, Niederegger H, Coultas L, Adams JM, Strasser A. Villunger A. BH3-only proteins Puma and Bim are rate limiting for gamma-radiation- and glucocorticoid-induced apoptosis of lymphoid cells in vivo. Blood. 106: 4131-4138, 2005.
[CrossRef] [PubMed]

Filderman RB, Kovacs BA. Anti-inflammatory activity of the steroid alkaloid glycoside, toatine. Br J Pharmacol. 37: 748-755, 1969.
[CrossRef] [PubMed]

Fontaine TD, Irwing GW, Ma R, Poole JB, Doolittle SP. Isolation and partial characterization of crystalline tomatine, an antibiotic agent from the tomato plant. Arch Biochem. 18: 467-475, 1948.
[PubMed]

Friedman M. Tomato glycoalkaloids: role in the plant and in the diet. J Agric Food Chem. 50: 5751-5780, 2002.
[CrossRef] [PubMed]

Friedman M, Levin CE, Lee SU, Kim HJ, Lee IS, Byun JO, Kozukue N. Tomatine-containing green tomato extracts inhibit growth of human breast, colon, liver, and stomach cancer cells. J Agric Food Chem. 57: 5727–5733, 2009.
[CrossRef] [PubMed]

Fuchs Y, Steller H. Programmed cell death in animal development and disease. Cell. 147: 742-758, 2011.
[CrossRef] [PubMed]

Irwing GW, Fontaine TD, Doolittle SP. Partial Antibiotic Spectrum of Tomatin, an Antibiotic Agent from the Tomato Plant. J Bacteriol. 52: 601-607, 1946.

Keukens EA, de Vrije T, van den Boom C, de Waard P, Plasman HH, Thiel F, Chupin V, Jongen WM, de Kruijff B. Molecular basis of glycoalkaloid induced membrane disruption. Biochim Biophys Acta. 1240: 216-228, 1995.
[CrossRef]

Keukens EA, de Vrije T, Jansen LA, de Boer H, Janssen M, de Kroon AI, Jongen WM, de Kruijff B. Glycoalkaloids selectively permeabilize cholesterol containing biomembranes. Biochim Biophys Acta. 1279: 243-250, 1996.
[CrossRef]

Lakin ND, Jackson SP. Regulation of p53 in response to DNA damage. Oncogene. 18: 7644-7655, 1999.
[CrossRef] [PubMed]

Lane DP. Cancer. P53, guardian of the genome. Nature. 358: 15-16, 1992.
[CrossRef] [PubMed]

Lavin MF, Gueven N. The complexity of p53 stabilization and activation. Cell Death Differ. 13: 941-950, 2006.
[CrossRef] [PubMed]

Lee KR, Kozukue N, Han JS, Park JH, Chang EY, Baek EJ, Chang JS, Friedman M. Glycoalkaloids and metabolites inhibit the growth of human colon (HT29) and liver (HepG2) cancer cells. J Agric Food Chem. 19: 2832-2839, 2004.
[CrossRef] [PubMed]

Lee ST, Wong PF, Cheah SC, Mustafa MR. Alpha-tomatine induces apoptosis and inhibits nuclear factor-kappa B activation on human prostatic adenocarcinoma PC-3 cells. PLoS One. 6: e18915, 2011.
[CrossRef] [PubMed]

Ma R, Fontaine TD. In vitro antibiotic activity of crystalline tomatine toward Candida albicans; antagonistic effect of rutin and quercetin. Arch Biochem. 16: 399-402, 1948.
[PubMed]

Olive PL, Banath JP. Detection of DNA double-strand breaks through the cell cycle after exposure to X-rays, bleomycin, etoposide and 125IdUrd. Int J Radiat Biol. 64: 349-358, 1993.
[CrossRef] [PubMed]

Prives C, Hall PA. The p53 pathway. J. Pathol. 187: 112–126, 1999.
[CrossRef]

Shi MD, Shih YW, Lee YS, Cheng YF, Tsai LY. Suppression of 12-O-Tetradecanoylphorbol-13-Acetate-Induced MCF-7 Breast Adenocarcinoma Cells Invasion/Migration by alpha-Tomatine Through Activating PKC?/ERK/NF-alphaB-Dependent MMP-2/MMP-9 Expressions. Cell Biochem Biophys. 2012 Nov 1. [Epub ahead of print].
[CrossRef]

Shieh JM, Cheng TH, Shi MD, Wu PF, Chen Y, Ko SC, Shih YW. Alpha-Tomatine Suppresses Invasion and Migration of Human Non-Small Cell Lung Cancer NCI-H460 Cells through Inactivating FAK/PI3K/Akt Signaling Pathway and Reducing Binding Activity of NF-kappa B. Cell Biochem Biophys. 60: 297-310, 2011.
[CrossRef] [PubMed]

Shieh SY, Ikeda M, Taya Y, Prives C. DNA damage induced phosporylation of p53 alleviates inhibition by MDM2. Cell. 91: 325-334, 1997.
[CrossRef]

Shih YW, Shieh JM, Wu PF, Lee YC, Chen YZ, Chiang TA. Alpha-tomatine inactivates PI3K/Akt and ERK signaling pathways in human lung adenocarcinoma A549 cells: effect on metastasis. Food Chem Toxicol. 47: 1985-1995, 2009.
[CrossRef] [PubMed]

Sionov RV, Haupt Y. The cellular response to p53: the decision between life and death. Oncogene. 18: 6145-6157, 1999.
[CrossRef] [PubMed]

Taylor WR, Stark GR. Regulation of the G2/M transition by p53. Oncogene. 20: 1803-1815, 2001.
[CrossRef] [PubMed]

Tichy A, Zaskodova D, Rezacova M, Vavrova J, Vokurkova D, Pejchal J, Vilasova Z, Cerman J, Osterreicher J. Gamma-radiation-induced ATM-dependent signalling in human T-lymphocyte leukemic cells, MOLT-4. Acta Biochim Pol. 54: 281-287, 2007.
[PubMed]

Wilson RH, Poley GW, De Eds F. Some pharmacologic and toxicologic properties of tomatine and its derivatives. Toxicol Appl Pharmacol. 3: 39-48, 1961.
[CrossRef]

Yap DBS, Hsieh J-K, Zhong S, Heath V, Gusterson B, Crook T, Lu X. Ser392 phosphorylation regulates the oncogenic function of mutant p53. Cancer Rest. 64: 4749–4754, 2004.
[CrossRef] [PubMed]
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