Effects of ghrelin, leptin and melatonin on the levels of reactive oxygen species, antioxidant enzyme activity and viability of the HCT 116 human colorectal carcinoma cell line

Bułdak,Rafał Jakub; Pilc‑Gumuła,Katarzyna; Bułdak,Łukasz; Witkowska,Daria; Kukla,Michał; Polaniak,Renata; Zwirska‑Korczala,Krystyna

doi:10.3892/mmr.2015.3599

Effects of ghrelin, leptin and melatonin on the levels of reactive oxygen species, antioxidant enzyme activity and viability of the HCT 116 human colorectal carcinoma cell line

Authors:
- Rafał Jakub Bułdak
- Katarzyna Pilc‑Gumuła
- Łukasz Bułdak
- Daria Witkowska
- Michał Kukla
- Renata Polaniak
- Krystyna Zwirska‑Korczala
View Affiliations

Affiliations: Department of Physiology, School of Medicine with the Division of Dentistry, Medical University of Silesia, Zabrze 41‑808, Poland, Department of Internal Medicine and Clinical Pharmacology, School of Medicine, Medical University of Silesia, Katowice 40‑752, Poland, Department of Gastroenterology and Hepatology, School of Medicine, Medical University of Silesia, Katowice 40‑752, Poland, Department of Human Nutrition, School of Public Health, Medical University of Silesia, Bytom 41‑902, Poland
Published online on: April 7, 2015 https://doi.org/10.3892/mmr.2015.3599
Pages: 2275-2282

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Obesity is associated with an increased risk of certain types of cancer, including colon cancer. Adipose tissue is an endocrine organ that produces biologically active substances, such as leptin and ghrelin. Recent research has suggested that adipose‑derived hormones may be associated with mechanisms linked to tumorigenesis and cancer progression. Furthermore, previous studies have demonstrated that pineal gland‑derived melatonin possesses important oncostatic and antioxidant properties. The present study aimed to determine the effects of the adipokines ghrelin and leptin, and the melatonin on intracellular levels of reactive oxygen species (ROS) and the activity of selected antioxidant enzymes, such as superoxide dismutase, catalase (CAT) and glutathione peroxidase. The effects of these compounds were also determined on the viability of HCT 116 human colorectal carcinoma cells in vitro. The pro‑oxidant and growth inhibitory effects of melatonin resulted in an accumulation of ROS and decreased antioxidant capacity in melatonin‑treated cells. Ghrelin administration alone caused a significant decrease in the levels of ROS, due to an increased activity of CAT in the HCT 116 cells. In addition, the present study observed increased lipid peroxidation following melatonin treatment, and decreased levels of malondialdehyde following ghrelin or leptin treatment. In conclusion, ghrelin, leptin and melatonin have various influences on the antioxidant capacity of HCT 116 cells. Compared with the adipokines, treatment with melatonin increased ROS levels and decreased cellular viability.

View Figures

View References

1	Housa D, Housová J, Vernerová Z and Haluzík M: Adipocytokines and cancer. Physiol Res. 55:233–244. 2006.
2	Pischon T, Nöthlings U and Boeing H: Obesity and cancer. Proc Nutr Soc. 67:128–145. 2008. View Article : Google Scholar : PubMed/NCBI
3	Mantovani G, Macciò A, Madeddu C, et al: Reactive oxygen species, antioxidant mechanisms and serum cytokine levels in cancer patients: Impact of an antioxidant treatment. J Cell Mol Med. 6:570–582. 2002. View Article : Google Scholar
4	Wiecek A, Kokot F, Chudek J and Adamczak M: The adipose tissue - a novel endocrine organ of interest to the nephrologist. Nephrol Dial Transplant. 17:191–195. 2002. View Article : Google Scholar : PubMed/NCBI
5	Kojima M, Hosoda H, Date Y, Nakazato M, Matsuo H and Kangawa K: Ghrelin is a growth-hormone-releasing acylated peptide from stomach. Nature. 402:656–660. 1999. View Article : Google Scholar : PubMed/NCBI
6	van der Lely AJ, Tschöp M, Heiman ML and Ghigo E: Biological, physiological, pathophysiological, and pharmacological aspects of ghrelin. Endocr Rev. 25:426–457. 2004. View Article : Google Scholar : PubMed/NCBI
7	Suzuki H, Matsuzaki J and Hibi T: Ghrelin and oxidative stress in gastrointestinal tract. J Clin Biochem Nutr. 48:122–125. 2011. View Article : Google Scholar : PubMed/NCBI
8	He X-T, Fan X-M and Zha X-L: Ghrelin inhibits 5-fluorouracil-induced apoptosis in colonic cancer cells. J Gastroenterol Hepatol. 26:1169–1173. 2011. View Article : Google Scholar : PubMed/NCBI
9	Nanzer AM, Khalaf S, Mozid AM, et al: Ghrelin exerts a proliferative effect on a rat pituitary somatotroph cell line via the mitogen-activated protein kinase pathway. Eur J Endocrinol. 151:233–240. 2004. View Article : Google Scholar : PubMed/NCBI
10	Gnanapavan S, Kola B, Bustin SA, et al: The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHS-R, in humans. J Clin Endocrinol Metab. 87:29882002. View Article : Google Scholar : PubMed/NCBI
11	Tian PY and Fan XM: The proliferative effects of ghrelin on human gastric cancer AGS cells. J Dig Dis. 13:453–458. 2012. View Article : Google Scholar : PubMed/NCBI
12	Barazzoni R, Zanetti M, Semolic A, Cattin MR, Pirulli A, Cattin L and Guarnieri G: High-fat diet with acyl-ghrelin treatment leads to weight gain with low inflammation, high oxidative capacity and normal triglycerides in rat muscle. PLoS One. 6:e262242011. View Article : Google Scholar : PubMed/NCBI
13	Konturek PC, Brzozowski T, Pajdo R, et al: Ghrelin-a new gastroprotective factor in gastric mucosa. J Physiol Pharmacol. 55:325–336. 2004.PubMed/NCBI
14	El Eter E, Al Tuwaijiri A, Hagar H and Arafa M: In vivo and in vitro antioxidant activity of ghrelin: Attenuation of gastric ischemic injury in the rat. J Gastroenterol Hepatol. 22:1791–1799. 2007. View Article : Google Scholar : PubMed/NCBI
15	Zhang Y, Proenca R, Maffei M, Barone M, Leopold L and Friedman JM: Positional cloning of the mouse obese gene and its human homologue. Nature. 372:425–432. 1994. View Article : Google Scholar : PubMed/NCBI
16	Aloulou N, Bastuji-Garin S, Le Gouvello S, et al: Involvement of the leptin receptor in the immune response in intestinal cancer. Cancer Res. 68:9413–9422. 2008. View Article : Google Scholar : PubMed/NCBI
17	Trayhurn P and Beattie JH: Physiological role of adipose tissue: White adipose tissue as an endocrine and secretory organ. Proc Nutr Soc. 60:329–339. 2001. View Article : Google Scholar : PubMed/NCBI
18	Watson AM, Poloyac SM, Howard G and Blouin RA: Effect of leptin on cytochrome P-450, conjugation, and antioxidant enzymes in the ob/ob mouse. Drug Metab Dispos. 27:695–700. 1999.PubMed/NCBI
19	Yuan Y, Zhang J, Cai L, et al: Leptin induces cell proliferation and reduces cell apoptosis by activating c-myc in cervical cancer. Oncol Rep. 29:2291–2296. 2013.PubMed/NCBI
20	Wu X, Yan Q, Zhang Z, Du G and Wan X: Acrp30 inhibits leptin-induced metastasis by downregulating the JAK/STAT3 pathway via AMPK activation in aggressive SPEC-2 endometrial cancer cells. Oncol Rep. 27:1488–1496. 2012.PubMed/NCBI
21	Yoon K-W, Park S-Y, Kim J-Y, et al: Leptin-induced adhesion and invasion in colorectal cancer cell lines. Oncol Rep. 31:2493–2498. 2014.PubMed/NCBI
22	Brydon L, Petit L, Delagrange P, Strosberg AD and Jockers R: Functional expression of MT2 (Mel1b) melatonin receptors in human PAZ6 adipocytes. Endocrinology. 142:4264–4271. 2001. View Article : Google Scholar : PubMed/NCBI
23	Reiter RJ: Melatonin: Lowering the high price of free radicals. News Physiol Sci. 15:246–250. 2000.
24	Baydaş G, Erçel E, Canatan H, Dönder E and Akyol A: Effect of melatonin on oxidative status of rat brain, liver and kidney tissues under constant light exposure. Cell Biochem Funct. 19:37–41. 2001. View Article : Google Scholar
25	Rodriguez C, Mayo JC, Sainz RM, Antolín I, Herrera F, Martín V and Reiter RJ: Regulation of antioxidant enzymes: A significant role for melatonin. J Pineal Res. 36:1–9. 2004. View Article : Google Scholar
26	Martín-Renedo J, Mauriz JL, Jorquera F, Ruiz-Andrés O, González P and González-Gallego J: Melatonin induces cell cycle arrest and apoptosis in hepatocarcinoma HepG2 cell line. J Pineal Res. 45:532–540. 2008. View Article : Google Scholar : PubMed/NCBI
27	Fan L-L, Sun G-P, Wei W, Wang Z-G, Ge L, Fu W-Z and Wang H: Melatonin and doxorubicin synergistically induce cell apoptosis in human hepatoma cell lines. World J Gastroenterol. 16:1473–1481. 2010. View Article : Google Scholar : PubMed/NCBI
28	Osseni RA, Rat P, Bogdan A, Warnet JM and Touitou Y: Evidence of prooxidant and antioxidant action of melatonin on human liver cell line HepG2. Life Sci. 68:387–399. 2000. View Article : Google Scholar
29	Petranka J, Baldwin W, Biermann J, Jayadev S, Barrett JC and Murphy E: The oncostatic action of melatonin in an ovarian carcinoma cell line. J Pineal Res. 26:129–136. 1999. View Article : Google Scholar : PubMed/NCBI
30	LeBel CP, Ischiropoulos H and Bondy SC: Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol. 5:227–231. 1992. View Article : Google Scholar : PubMed/NCBI
31	Bułdak RJ, Polaniak R, Bułdak L, et al: Short-term exposure to 50 Hz ELF-EMF alters the cisplatin-induced oxidative response in AT478 murine squamous cell carcinoma cells. Bioelectromagnetics. 33:641–651. 2012. View Article : Google Scholar
32	Paglia DE and Valentine WN: Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 70:158–169. 1967.PubMed/NCBI
33	Bułdak RJ, Bułdak Ł, Polaniak R, et al: Visfatin affects redox adaptative responses and proliferation in Me45 human malignant melanoma cells: An in vitro study. Oncol Rep. 29:771–778. 2013.
34	Paoletti F and Mocali A: Determination of superoxide dismutase activity by purely chemical system based on NAD(P)H oxidation. Methods Enzymol. 186:209–220. 1990.PubMed/NCBI
35	Aebi H: Catalase in vitro. Methods Enzymol. 105:121–126. 1984.PubMed/NCBI
36	Ohkawa H, Ohishi N and Yagi K: Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 95:351–358. 1979. View Article : Google Scholar : PubMed/NCBI
37	Anisimov VN, Popovich IG, Zabezhinski MA, Anisimov SV, Vesnushkin GM and Vinogradova IA: Melatonin as antioxidant, geroprotector and anticarcinogen. Biochim Biophys Acta. 1757:573–589. 2006. View Article : Google Scholar : PubMed/NCBI
38	Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M and Telser J: Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 39:44–84. 2007. View Article : Google Scholar
39	Koda M, Sulkowska M, Kanczuga-Koda L, Surmacz E and Sulkowski S: Overexpression of the obesity hormone leptin in human colorectal cancer. J Clin Pathol. 60:902–906. 2007. View Article : Google Scholar : PubMed/NCBI
40	Aparicio T, Kotelevets L, Tsocas A, et al: Leptin stimulates the proliferation of human colon cancer cells in vitro but does not promote the growth of colon cancer xenografts in nude mice or intestinal tumorigenesis in Apc(Min/+) mice. Gut. 54:1136–1145. 2005. View Article : Google Scholar : PubMed/NCBI
41	Zhou J, Lei W, Shen L, Luo H-S and Shen Z-X: Primary study of leptin and human hepatocellular carcinoma in vitro. World J Gastroenterol. 14:2900–2904. 2008. View Article : Google Scholar : PubMed/NCBI
42	Catalano S, Marsico S, Giordano C, et al: Leptin enhances, via AP-1, expression of aromatase in the MCF-7 cell line. J Biol Chem. 278:28668–28676. 2003. View Article : Google Scholar : PubMed/NCBI
43	Chang S, Hursting SD, Contois JH, et al: Leptin and prostate cancer. Prostate. 46:62–67. 2001. View Article : Google Scholar : PubMed/NCBI
44	Gonzalez RR, Cherfils S, Escobar M, et al: Leptin signaling promotes the growth of mammary tumors and increases the expression of vascular endothelial growth factor (VEGF) and its receptor type two (VEGF-R2). J Biol Chem. 281:26320–26328. 2006. View Article : Google Scholar : PubMed/NCBI
45	Yamagishi SI, Edelstein D, Du XL, Kaneda Y, Guzmán M and Brownlee M: Leptin induces mitochondrial superoxide production and monocyte chemoattractant protein-1 expression in aortic endothelial cells by increasing fatty acid oxidation via protein kinase A. J Biol Chem. 276:25096–25100. 2001. View Article : Google Scholar : PubMed/NCBI
46	Xu F-P, Chen M-S, Wang Y-Z, Yi Q, Lin S-B, Chen AF and Luo J-D: Leptin induces hypertrophy via endothelin-1-reactive oxygen species pathway in cultured neonatal rat cardiomyocytes. Circulation. 110:1269–1275. 2004. View Article : Google Scholar : PubMed/NCBI
47	Zwirska-Korczala K, Adamczyk-Sowa M, Sowa P, et al: Role of leptin, ghrelin, angiotensin II and orexins in 3T3 L1 preadipocyte cells proliferation and oxidative metabolism. J Physiol Pharmacol. 58(Suppl 1): 53–64. 2007.PubMed/NCBI
48	Balasubramaniyan V, Shukla R, Murugaiyan G, Bhonde RR and Nalini N: Mouse recombinant leptin protects human hepatoma HepG2 against apoptosis, TNF-alpha response and oxidative stress induced by the hepatotoxin-ethanol. Biochim Biophys Acta. 1770:1136–1144. 2007. View Article : Google Scholar : PubMed/NCBI
49	Li WG, Gavrila D, Liu X, et al: Ghrelin inhibits proinflammatory responses and nuclear factor-kappaB activation in human endothelial cells. Circulation. 109:2221–2226. 2004. View Article : Google Scholar : PubMed/NCBI
50	Valko M, Rhodes CJ, Moncol J, Izakovic M and Mazur M: Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact. 160:1–40. 2006. View Article : Google Scholar : PubMed/NCBI
51	Tong X-X, Wu D, Wang X, et al: Ghrelin protects against cobalt chloride-induced hypoxic injury in cardiac H9c2 cells by inhibiting oxidative stress and inducing autophagy. Peptides. 38:217–227. 2012. View Article : Google Scholar : PubMed/NCBI
52	Kheradmand A, Alirezaei M and Birjandi M: Ghrelin promotes antioxidant enzyme activity and reduces lipid peroxidation in the rat ovary. Regul Pept. 162:84–89. 2010. View Article : Google Scholar : PubMed/NCBI
53	Roth JA, Rosenblatt T, Lis A and Bucelli R: Melatonin-induced suppression of PC12 cell growth is mediated by its Gi coupled transmembrane receptors. Brain Res. 919:139–146. 2001. View Article : Google Scholar : PubMed/NCBI
54	Lamosová D, Zeman M and Juráni M: Influence of melatonin on chick skeletal muscle cell growth. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 118:375–379. 1997. View Article : Google Scholar
55	Chen LD, Leal BZ, Reiter RJ, et al: Melatonin’s inhibitory effect on growth of ME-180 human cervical cancer cells is not related to intracellular glutathione concentrations. Cancer Lett. 91:153–159. 1995. View Article : Google Scholar : PubMed/NCBI
56	Cini G, Coronnello M, Mini E and Neri B: Melatonin’s growth-inhibitory effect on hepatoma AH 130 in the rat. Cancer Lett. 125:51–59. 1998. View Article : Google Scholar : PubMed/NCBI
57	Roberts JE, Wiechmann AF and Hu DN: Melatonin receptors in human uveal melanocytes and melanoma cells. J Pineal Res. 28:165–171. 2000. View Article : Google Scholar : PubMed/NCBI
58	Gilad E, Laudon M, Matzkin H, et al: Functional melatonin receptors in human prostate epithelial cells. Endocrinology. 137:1412–1417. 1996.PubMed/NCBI
59	Lupowitz Z and Zisapel N: Hormonal interactions in human prostate tumor LNCaP cells. J Steroid Biochem Mol Biol. 68:83–88. 1999. View Article : Google Scholar : PubMed/NCBI
60	Dziegiel P, Podhorska-Okolow M, Surowiak P, Ciesielska U, Rabczynski J and Zabel M: Influence of exogenous melatonin on doxorubicin-evoked effects in myocardium and in transplantable Morris hepatoma in rats. In Vivo. 17:325–328. 2003.PubMed/NCBI
61	Pirozhok I, Meye A, Hakenberg OW, Fuessel S and Wirth MP: Serotonin and melatonin do not play a prominent role in the growth of prostate cancer cell lines. Urol Int. 84:452–460. 2010. View Article : Google Scholar : PubMed/NCBI
62	Jang SS, Kim WD and Park W-Y: Melatonin exerts differential actions on X-ray radiation-induced apoptosis in normal mice splenocytes and Jurkat leukemia cells. J Pineal Res. 47:147–155. 2009. View Article : Google Scholar : PubMed/NCBI
63	Kim J-H, Jeong S-J, Kim B, Yun S-M, Choi Y and Kim S-H: Melatonin synergistically enhances cisplatin-induced apoptosis via the dephosphorylation of ERK/p90 ribosomal S6 kinase/heat shock protein 27 in SK-OV-3 cells. J Pineal Res. 52:244–252. 2012. View Article : Google Scholar
64	Lamson DW and Brignall MS: Antioxidants in cancer therapy; their actions and interactions with oncologic therapies. Altern Med Rev. 4:304–329. 1999.PubMed/NCBI
65	Polaniak R, Bułdak RJ, Karoń M, et al: Influence of an extremely low frequency magnetic field (ELF-EMF) on antioxidative vitamin E properties in AT478 murine squamous cell carcinoma culture in vitro. Int J Toxicol. 29:221–230. 2010. View Article : Google Scholar : PubMed/NCBI