Hande CANPINAR, Şükran YILMAZ, Güldeniz SELMANOĞLU, Aysun SÜLOĞLU KILIÇ

Comparison of phototoxic effects of hypericin-mediated photodynamic therapy in HT-29 and Caco-2 colon cancer cells

Hypericin (HYP) is a plant-derived photosensitizer. HYP is preferentially taken up by tumor cells. We designed this study to compare HYP-mediated photodynamic therapy (PDT) in HT-29 and Caco-2 colon cell lines. Cells were treated with 0.04, 0.08, and 0.15 μM HYP concentrations and irradiated. The effect of HYP on metabolic profiles, alterations in lactate dehydrogenase (LDH) leakage, and cell cycle progression was investigated for the first time. Changes in glucose consumption, lactate production, and LDH leakage were analyzed. HYP-induced cell death was quantified by double staining (acridine orange/propidium iodide) and alterations in cell cycle regulation were analyzed with flow cytometry (using propidium iodide). LDH leakage and the number of dead cells were elevated, and glucose consumption and lactate production decreased in a dose- and time-dependent manner. PDT resulted in an induction of apoptosis, mostly at the 0.08 μM HYP concentration. Apoptosis and/or necrosis were increased in the 0.15 μM HYP group. The accumulation of cells in the G2/M phase might account for the growth inhibition in HT-29 and Caco-2 cells with 0.08 μM HYP photoactivation. The observed G2/M arrest suggested that HYP may slow down the growth of colon cancer cells by regulating the cell cycle, leading either to growth inhibition or to initiation of apoptotic pathways.

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American Cancer Society (2011). Colorectal Cancer Facts and Figures. Atlanta, GA, USA: American Cancer Society.Barathan M, Mariappan V, Shankar EM, Abdullah BJJ, Goh KL, Vadivelu J (2013). Hypericin-photodynamic therapy leads to interleukin-6 secretion by HEPG2 cells and their apoptosis via recruitment of BH-3 interacting-domain death agonist and caspases. Cell Death Dis 27: e697.Boya P, González-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N, Métivier D, Meley D, Souquere S, Yoshimori T et al. (2005). Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol 25: 1025-1040.Dang CV, Semenza GL (1999). Oncogenic alterations of metabolism. Trends Biochem Sci 24: 68-72.DiPaola RS (2002). To arrest or not to G2-M cell cycle arrest. Clin Cancer Res 8: 3311-3314.Foglieni C, Meoni C, Davalli AM (2001). Fluorescent dyes for cell viability: an application on prefixed conditions. Histochem Cell Biol 115: 223-229.Gatenby RA, Gillies RJ (2004). Why do cancers have high aerobic glycolysis? Nat Rev Cancer 4: 891-899.
Hendrickx N, Volanti C, Moens U, Seternes OM, Witte P, Vandenheede JR, Piette J, Agostinis P (2003). Up-regulation of cyclooxygenase-2 and apoptosis resistance by p38 MAPK in hypericin-mediated photodynamic therapy of human cancer cells. J Biol Chem 278: 52231-52239.Hoiroyde CP, Axelrod RS, Skutches CL, Haff AC, Paul P, Reichard GA (1979). Lactate metabolism in patients with metastatic colorectal cancer. Cancer Res 39: 4900-4904.Johnson SA, Dalton AE, Pardini RS (1998). Time-course of hypericin phototoxicity and effect on mitochondrial energies in EMT6 mouse mammary carcinoma cells. Free Radical Bio Med 25: 144-152.Karioti A, Bilia AR (2010). Hypericins as potential leads for new therapeutics. Int J Mol Sci 11: 562-594.Kiesslich T, Plaetzer K, Oberdanner CB, Berlanda J, Obermair FJ, Krammer B (2005). Differential effects of glucose deprivation on the cellular sensitivity towards photodynamic treatment-based production of reactive oxygen species and apoptosis-induction. FEBS Lett 579: 185-190.Kılıç Süloğlu A, Karacaoğlu E, Selmanoğlu G, Akel H, Karaaslan İÇ (2016). Evaluation of apoptotic cell death mechanisms induced by hypericin mediated photodynamic therapy in colon cancer cells. Turk J Biol 40: 539-546.Kılıç Süloğlu A, Selmanoğlu G, Akay MT (2015). Alterations in dysadherin expression and F-actin reorganization: a possible mechanism of hypericin mediated photodynamic therapy in colon adenocarcinoma cells. Cytotechnology 67: 311-330.Kleban J, Mike J, Horváth V, Sačková V, Hofmanová J, Kozubík A, Fedoročko P (2008). Mechanisms involved in the cell cycle and apoptosis of HT-29 cells pre-treated with MK-886 prior to photodynamic therapy with hypericin. J Photoch Photobio B 93: 108-118.Legrand C, Bour JM, Jacob C, Capiaumont J, Martial A, Marc A, Wudtke M, Kretzmer G, Demangel C, Duval D et al. (1992). Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker. J Biotechnol 25: 231-243.Lev-Ari S, Strier L, Kazanov D, Shapiro LM, Sobol HD, Pinchuk I, Marian B, Lichtenberg D, Arber N (2005). Celecoxib and curcumin synergistically inhibit the growth of colorectal cancer cells. Clin Cancer Res 11: 6738-6744.Nihal A, Feyes DK, Nieminen AL, Agarwal R, Mukhtar H (1997). Green tea constituent epigallocatechin-3-gallate and induction of apoptosis and cell cycle arrest in human carcinoma cells. J Nat Cancer Ins 89: 24.Riccardi C, Nicoletti I (2006). Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1: 1458-1461.Sačková V, Fedoročko P, Szllardiová B, Mike J, Kleban J (2006). Hypericin-induced photocytotoxicity is connected with G2/M arrest in HT-29 and S-phase arrest in U937 cells. J Photoch Photobio B 82: 1285-1291.Sanovic R, Verwanger T, Hartl A, Krammer B (2011). Low dose hypericin-PDT induces complete tumor regression in BALB/c mice bearing CT26 colon carcinoma. Photodiagn Photodyn Ther 8: 291-296.Wang W, Heideman L, Chung CS, Pelling JC, Koehler KJ, Birt DF (2000). Cell-cycle arrest at G2/M and growth inhibition by apigenin in human colon carcinoma cell lines. Mol Carcinogen 28: 102-110.Yang ZR, Liu M, Peng XL, Lei XF, Zhang JX, Dong WG (2012). Noscapine induces mitochondria-mediated apoptosis in human colon cancer cells in vivo and in vitro. Biochem Bioph Res Co 421: 627-633.Young DS (1995). Effects of Drugs on Clinical Laboratory Tests. 4th ed. Washington, DC, USA: AACC Press.