Advances in Clinical and Experimental Medicine
2015, vol. 24, nr 1, January-February, p. 37–46
Publication type: original article
Expression of Proapoptotic BAX and TP53 Genes and Antiapoptotic BCL-2 Gene in MCF-7 and T-47D Tumour Cell Cultures of the Mammary Gland After a Photodynamic Therapy with Photolon
1 Department of Cell Biology, School of Pharmacy, Medical University of Silesia, Sosnowiec, Poland
Background. Breast cancer is the most common malignant tumour in women in the whole world. Despite significant developments in the early diagnosis of breast cancer, there is no effective method which would assure total recovery of the patient. Currently available clinical data and laboratory tests indicate a possibility to introduce photodynamic therapy (PDT) to the supplementary treatment of breast cancer.
Objectives. The aim of this study was to assess the influence of PDT with Photolon as a photosensibilizator on the expression of apoptosis associated genes (BCL-2, BAX, TP53) in human breast cancer cell lines, preceded by assessment of survivorship and proliferative activity in the tested cells after PDT.
Material and Methods. In the present study human breast cancer cell lines MCF-7 and T-47D were used. Photolon (chlorin e6 complex: PVP 1:1) was used as a photosensitizer. Assessments of survivorship and proliferative activity of cells under the influence of PDT (WST-1 test) were conducted along with the expression of selected genes involved in the process of apoptosis: BCL-2, BAX, TP53 (RT-QPCR).
Results. PDT limited both survivorship and proliferative activity of breast cancer cells in the two tested lines. In case of T-47D cell line was found increase of BAX and BCL-2 genes expression after PDT and sustained activity of TP53 gene. Conversely, in MCF-7 cell line a decrease in expression was found for both BAX and TP53 genes, but also an increase of BCL-2 gene expression.
Conclusion. A progressing decrease (24, 48 and 72 h after PDT) in the count of culture cells, which suggests the occurrence of apoptosis initiated by a photodynamic reaction with simultaneous increase of BCL-2/BAX index, indicates activation of a different endogenous apoptosis pathway than the one examined, namely pointing to suicidal death of cells after PDT.
photodynamic therapy, Photolon, MCF-7, T-47D, apoptosis
- Jassem J, Krzakowski M: Rak piersi. Via Medica, Gdańsk 2009.
- Allison RR, Sibata C, Downie GH, Cuenca RE: Photodynamic therapy of the intact breast. Photodiag Photodyn Ther 2006, 3, 139–146.
- Pruthi S, Brand KR, Degnim AC, Goetz MP, Perez EA, Reynolds CA, Schomberg PJ, Dy GK: A multidisciplinary approach to the management of breast cancer, part 1: prevention and diagnosis. Mayo Clin Proc 2007, 82, 999–1012.
- Pruthi S, Boughey JC, Brandt KR, Degnim AC, Dy GK, Goetz MP, Perez EA, Reynolds CA, Schomberg PJ, Ingle JN: A multidisciplinary approach to the management of breast cancer, part 2: therapeutic considerations. Mayo Clin Proc 2007, 82, 1131–1140.
- Robertson CA, Hawkins Evans D, Abrahamse H: Photodynamic therapy (PDT): A short review on cellular mechanisms and cancer research applications for PDT. J Photochem Photobiol B 2009, 96, 1–8.
- Nowak-Stępniowska A, Małecki M, Wiktorska K, Romiszewska A, Padzik-Graczyk A: Inhibition of cell growth induced by photosensitizer PP(Arg)2-mediated photodynamic therapy in human breast and prostate cell lines. Part I. Photodiag Photodyn Ther 2011, 8, 39–48.
- Mroz P, Yaroslavsky A, Kharkwal GB, Hamblin MR: Cell death pathways in photodynamic therapy of cancer. Cancer 2011, 3, 2516–2539.
- Buytaert E, Dewaele M, Agostinis P: Molecular effectors of multiple cell death pathways initiated by photodynamic therapy. Biochim Biophys Acta 2007, 1776, 86–107.
- Montazerabadi AR, Sazgarnia A, Bahreyni-Toosi MH, Ahmadi A, Shakeri-Zadeh A, Aledavood A: Mitoxantrone as a prospective photosensitizer for photodynamic therapy of breast cancer. Photodiag Photodyn Ther 2012, 9, 46–51.
- Wezgowiec J, Kotulska M, Saczko J, Derylo MB, Teissie J, Rols MP, Orio J, Garbiec A, Kulbacka J: Cyanines in photodynamic reaction assisted by reversible electroporation – in vitro study on human breast carcinoma cells. Photodiag Photodyn Ther 2013, 10, 490–502.
- Li B, Chu X, Gao M, Li W: Apoptotic mechanism of MCF-7 breast cells in vivo and in vitro induced by photodynamic therapy with C-phycocyanin. Acta Biochim Biophys Sin 2010, 42, 80–89.
- Wyld L, Reed MWR, Brown NJ: Differential cell death response to photodynamic therapy is dependent on dose and cell type. Br J Cancer 2001, 84, 1384–1386.
- Marchal S, Francois A, Dumas D, Guillemin F, Bezdetnaya L: Relationship between subcellular localization of Foscan® and caspase activation in photosensitized MCF-7 cells. Br J Cancer 2007, 96, 944–951.
- Misiewicz-Krzemińska I, Skupińska K, Graczyk A, Kasprzycka-Guttman T: Influence of protoporphyrin IX amino acid substituents on affinity to human breast adenocarcinoma MCF-7 cells. Biotech Histochem 2009, 84, 17–23.
- Schlotter CM, Vogt U, Allgayer H, Brandt B: Molecular targeted therapies for breast cancer treatment. Breast Cancer Res 2008, 10, 211.
- Koval J, Mikes J, Jendzelovsky R, Kello M, Solar P, Fedorocko P: Degradation of HER 2 receptor through hypericinmediated photodynamic therapy. Photochem Photobiol 2010, 86, 200–205.
- Xue L, Chiu S, Fiebig A, Andrews DW, Oleinick NL: Photodamage to multiple Bcl-xL isoforms by photodynamic therapy with the phthalocyanine photosensitizer Pc 4. Oncogene 2003, 22, 9197–9204.
- Copley L, van der Watt P, Wirtz KW, Parker MI, Leaner VD: PhotolonTM, a chlorine derivative, triggers ROS production and light-dependent cell death via necrosis. Int J Biochem Cell Biol 2008, 40, 227–235.
- Plaetzer K, Krammer B, Berlanda J, Berr F, Kiesslich T: Photophysics and photochemistry of photodynamic therapy: fundamental aspects. Lasers Med Sci 2009, 24, 259–268.
- Burch S, Bisland SK, Bogaards A, Yee AJ, Whyne CM, Finkelstein JA, Wilson BC: Photodynamic therapy for the treatment of vertebral metastases in a rat model of human breast carcinoma. J Orthop Res 2005, 23, 995–1003.
- Allison R, Mang T, Hewson G, Snider W, Dougherty D: Photodynamic therapy for chest wall progression from breast carcinoma is an underutilized treatment modality. Cancer 2001, 91, 1–8.
- Jerjes W, Upile T, Mosse ChA, Hamdoon Z, Morcos M, Morley S, Hopper C: Prospective evaluation of 110 patients following ultrasound-guided photodynamic therapy for deep seated pathologies. Photodiag Photodyn Ther 2011, 8, 297–306.
- Lee CHL, Blum JM, Kirsch DG: Role of p53 in regulating tissue response to radiation by mechanisms independent of apoptosis. Transl Cancer Res 2013, 2, 412–421.
- Zawacka-Pankau J, Issaeva N, Hossain S, Pramanik A, Selivanova G, Podhajska AJ: Protoporphyrin IX interacts with wild-type p53 protein in vitro and induces cell death of human colon cancer cells in a p 53-dependent and – independent manner. J Biol Chem 2007, 282, 2466–2472.
- Lee HB, Ho ASH, Teo SH: P53 Status does not affect photodynamic cell killing induced by hypericin. Cancer Chemother Pharmacol 2006, 58, 91–98.
- Peltonen K, Kiviharju TM, Jarvinen PM, Ra R, Laiho M: Melanoma cell lines are susceptible to histone deacetylase inhibitor TSA provoked cell cycle arrest and apoptosis. Pigment Cell Res 2005, 18, 196–202.
- Chin WWL, Heng PWS, Olivo M: Chlorin e6 – polyvinylpyrrolidone mediated photosensitization is effective against human non-small cell lung carcinoma compared to small cell lung carcinoma xenografts. BMC Pharmacol [Internet] 2007, 7, 1–9. Available from: http://www.biomedcentral.com/ 1471-2210/ 7/ 15.
- Copley L, van der Watt P, Wirtz KW, Parker MI, Leaner VD: Photolon, a chlorin e6 derivative, triggers ROS production and light-dependent cell death via necrosis. IJBCB 2008, 40, 227–235.