Epigallocatechin 3-gallate applications on HT-29 and MCF-7 cell lines andevaluation of tumor suppressor gene methylation

Abstract: Epigallocatechin 3-gallate (EGCG) is an antitumor molecule and shows this activity by binding to the active center of a methyltransferase enzyme (DNMT1). The methylation of DNA sequences of tumor suppressor and DNA repair genes is observed in different stages of carcinogenesis. In this study, we analyzed the effect of EGCG on the methylation status of 25 tumor suppressor genes in cancer cell lines HT-29 and MCF-7. HT-29 and MCF-7 cells were incubated with 10 μM, 20 μM, and 50 μM and 1 μM, 5 μM, and 10 μM EGCG for 48 h, respectively. We found promoter hypermethylation of (1) CDH13, GATA5, and RARβ genes in MCF-7 cell line and (2) RARβ, ESR1, PAX6, WT1, CADM1, CHFR, CDH13, and GATA5 genes in HT-29 cell line. However, (3) after EGCG application, no changes in methylation status were detected in our samples. Our results suggest that methylation status of tumor suppressor genes did not change with different EGCG doses.

Epigallocatechin 3-gallate applications on HT-29 and MCF-7 cell lines andevaluation of tumor suppressor gene methylation

Abstract: Epigallocatechin 3-gallate (EGCG) is an antitumor molecule and shows this activity by binding to the active center of a methyltransferase enzyme (DNMT1). The methylation of DNA sequences of tumor suppressor and DNA repair genes is observed in different stages of carcinogenesis. In this study, we analyzed the effect of EGCG on the methylation status of 25 tumor suppressor genes in cancer cell lines HT-29 and MCF-7. HT-29 and MCF-7 cells were incubated with 10 μM, 20 μM, and 50 μM and 1 μM, 5 μM, and 10 μM EGCG for 48 h, respectively. We found promoter hypermethylation of (1) CDH13, GATA5, and RARβ genes in MCF-7 cell line and (2) RARβ, ESR1, PAX6, WT1, CADM1, CHFR, CDH13, and GATA5 genes in HT-29 cell line. However, (3) after EGCG application, no changes in methylation status were detected in our samples. Our results suggest that methylation status of tumor suppressor genes did not change with different EGCG doses.

___

  • Alvarez M, Robey R, Sandor V, Nishiyama K, Matsumoto Y, Paull K, Bates S, Fojo T (1998). Using the National Cancer Institute anticancer drug screen to assess the effect of MRP expression on drug sensitivity profiles. Mol Pharmacol 54: 802–814.
  • Arai M, Yokosuka O, Hirasawa Y, Fukai K, Chiba T, Imazeki F, Kanda T, Yatomi M, Takiguchi Y, Seki N et al. (2006). Sequential gene expression changes in cancer cell lines after treatment with the demethylation agent 5-Aza-2’-deoxycytidine. Cancer 106: 2514–2525.
  • Arends MJ (2013). Pathways of colorectal carcinogenesis. Appl Immunohistochem Mol Morphol 21: 97–102.
  • Baylin SB, Herman JG (2000). DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trends Genet 16: 168–174.
  • Brooks J, Cairns P, Zeleniuch-Jacquotte A (2009). Promoter methylation and the detection of breast cancer. Cancer Causes Control 20: 1539–1550.
  • Chen D, Wan SB, Yang H, Yuan J, Chan TH, Dou QP (2011). EGCG, green tea polyphenols and their synthetic analogs and prodrugs for human cancer prevention and treatment. Adv Clin Chem 53: 155–177.
  • Cho YH, Yazici H, Wu HC, Terry MB, Gonzalez K, Qu M, Dalay N, Santella RM (2010). Aberrant promoter hypermethylation and genomic hypomethylation in tumor, adjacent normal tissues and blood from breast cancer patients. Anticancer Res 30: 2489–2496.
  • Chung J Y, Huang C, Meng X, Dong Z, Yang CS (1999). Inhibition of activator protein 1 activity and cell growth by purified green tea and black tea polyphenols in H-Ras-transformed cells: structure-activity relationship and mechanisms involved. Cancer Res 59: 4610–4617.
  • Chung JY, Park JO, Phyu H, Dong Z, Yang CS (2001). Mechanisms of inhibition of the Ras-MAP kinase signaling pathway in 30.7b Ras 12 cells by tea polyphenols (-)-epigallocatechin-3-gallate and theaflavin-3,3’-digallate. FASEB J 15: 2022–2024.
  • Colussi D, Brandi G, Bazzoli F, Ricciardiello L (2013). Molecular pathways involved in colorectal cancer: implications for disease behavior and prevention. Int J Mol Sci 14: 16365–16385.
  • Fang MZ, Wang Y, Ai N, Hou Z, Sun Y, Lu H, Welsh W, Yang CS (2003). Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res 63: 7563–7570.
  • Gao Z, Xu Z, Hung MS, Lin YC, Wang T, Gong M, Zhi X, Jablon DM, You L (2009). Promoter demethylation of WIF-1 by epigallocatechin-3-gallate in lung cancer cells. Anticancer Res 29: 2025–2230.
  • Hardy TM, Tollefsbol TO (2011). Epigenetic diet: impact on the epigenome and cancer. Epigenomics 3: 503–518.
  • Hong J, Lu H, Meng X, Ryu JH, Hara Y, Yang CS (2002). Stability, cellular uptake, biotransformation, and efflux of tea polyphenol (-)-epigallocatechin-3-gallate in HT-29 human colon adenocarcinoma cells. Cancer Res 62: 7241–7246.
  • Hsu YC, Liou YM (2011). The anti-cancer effects of (-)-epigallocatechin-3-gallate on the signaling pathways associated with membrane receptors in MCF-7 cells. J Cell Physiol 226: 2721–2730.
  • Kars MD, Iseri OD, Gündüz U, Ural AU, Arpaci F, Molnár J (2006). Development of rational in vitro models for drug resistance in breast cancer and modulation of MDR by selected compounds. Anticancer Res 26: 4559–4568.
  • Kornegoor R, Moelans CB, Verschuur-Maes AH, Hogenes MCh, de Bruin PC, Oudejans JJ, van Diest PJ (2012). Promoter hypermethylation in male breast cancer: analysis by multiplex ligation-dependent probe amplification. Breast Cancer Res 14: R101.
  • Lecumberri E, Dupertuis YM, Miralbell R, Pichard C (2014). Green tea polyphenol epigallocatechin-3-gallate (EGCG) as adjuvant in cancer therapy. Clin Nutr 32: 894–903.
  • Lee WJ, Shim JY, Zhu BT (2005). Mechanisms for the inhibition of DNA methyltransferases by tea catechins and bioflavonoids. Mol Pharmacol 68: 1018–1030.
  • Li Y, Tollefsbol TO (2010). Impact on DNA methylation in cancer prevention and therapy by bioactive dietary components. Curr Med Chem 17: 2141–2151.
  • Liang YC, Lin-shiau SY, Chen CF, Lin JK (1997). Suppression of extracellular signals and cell proliferation through EGF receptor binding by (-)-epigallocatechin gallate in human A431 epidermoid carcinoma cells. J Cell Biochem 67: 55–65.
  • Moelans CB, Verschuur-Maes AH, van Diest PJ (2011). Frequent promoter hypermethylation of BRCA2, CDH13, MSH6, PAX5, PAX6 and WT1 in ductal carcinoma in situ and invasive breast cancer. J Pathol 225: 222–231.
  • Moison C, Senamaud-Beaufort C, Fourrière L, Champion C, Ceccaldi A, Lacomme S, Daunay A, Tost J, Arimondo PB, Guieysse-Peugeot AL (2013). DNA methylation associated with polycomb repression in retinoic acid receptor β silencing. FASEB J 27: 1468–1478.
  • Nandakumar V, Vaid M, Katiyar SK (2011). (-)-Epigallocatechin-3- gallate reactivates silenced tumor suppressor genes, Cip1/p21 and p16INK4a, by reducing DNA methylation and increasing histones acetylation in human skin cancer cells. Carcinogenesis 32: 537–544.
  • Nystrom M, Mutanen M (2009). Diet and epigenetics in colon cancer. World J Gastroenterol 15: 257–263.
  • Pandey M, Shukla S, Gupta S (2010). Promoter demethylation and chromatin remodeling by green tea polyphenols leads to re- expression of GSTP1 in human prostate cancer cells. Int J Cancer 126: 2520–33.
  • Park SY, Kwon HJ, Lee HE, Ryu HS, Kim SW, Kim JH, Kim IA, Jung N, Cho NY, Kang GH (2011). Promoter CpG island hypermethylation during breast cancer progression. Virchows Arch 458: 73–84.
  • Sachinidis A, Seul C, Seewald S, Ahn H, Ko Y, Vetter H (2000). Green tea compounds inhibit tyrosine phosphorylation of PDGF b-receptor and transformation of A172 human glioblastoma. FEBS Lett 471: 51–55.
  • Singh PN, Fraser GE (1998). Dietary risk factors for colon cancer in a low-risk population. Am J Epidemiol 148: 761–774.
  • Xiang TX, Yuan Y, Li LL, Wang ZH, Dan LY, Chen Y, Ren GS, Tao Q (2013). Aberrant promoter CpG methylation and its translational applications in breast cancer. Chin J Cancer 32: 12–20.
  • Xu J, Shetty PB, Feng W, Chenault C, Bast RC Jr, Issa JP, Hilsenbeck SG, Yu Y (2012). Methylation of HIN-1, RASSF1A, RIL and CDH13 in breast cancer is associated with clinical characteristics, but only RASSF1A methylation is associated with outcome. BMC Cancer 12: 243.
  • Zeng L, Holly JMP, Perks CM (2014). Effects of physiological levels of the green tea extract epigallocatechin-3-gallate on breast cancer cells. Front Endocrinol (Lausanne) 5: 61.
  • Zhang X, Min KW, Wimalasena J, Baek SJ (2012). Cyclin D1 degradation and p21 induction contribute to growth inhibition of colorectal cancer cells induced by epigallocatechin-3-gallate. J Cancer Res Clin Oncol 138: 2051–2060.
  • Zheng R, Blobel GA (2010). GATA transcription factors and cancer. Genes Cancer 1: 1178–1188.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

CDK inhibitors-induced SSAT expression requires NF?B and PPAR? inMCF-7 breast cancer cells

PINAR OBAKAN, ŞEYMA YILDIRIM, MERT BURAK ÖZTÜRK, ÖZGE BERRAK, AJDA ÇOKER GÜRKAN, ELİF DAMLA ARISAN, ZEYNEP NARÇİN ÜNSAL

Marker-assisted selection for rice brown planthopper (Nilaparvata lugens)resistance using linked SSR markers

MAHMOODREZA SHABANIMOFRAD, MOHD RAFII YUSOP, SADEGH ASHKANI, MOHAMED HANAFI MUSA, NUR AZURA ADAM, IFFAH HAIFA, ABDUL RAHIM HARUN, MOHAMMAD ABDUL LATIF

Immunohistochemical identification of aquaporin 2 in the kidneys of wild boars (Sus scrofa)

KATARZYNA MICHALEK, EWA CZERNIAWSKA-PIATKOWSKA, MARTA GRABOWSKA, MARIA LASZCZYNSKA

Effects of squirting cucumber (Ecballium elaterium) fruit juice onAgrobacterium tumefaciens-mediated transformation of plants

SANCAR FATİH ÖZCAN, MUSTAFA YILDIZ, HUSSEIN ABDULLAH AHMED AHMED, MUHAMMAD AASIM

Improvement of seed germination and in vitro propagation of Bupleurum latissimum Nakai through embryogenesis

KEE HWA BAE

Discovery of Dhn3 variants in wild barley (Hordeum spontaneum) byhigh-resolution melting (HRM) technology

CÜNEYT UÇARLI, NAZARET POYRAZ, AYŞE FİLİZ GÜREL

Epigallocatechin 3-gallate applications on HT-29 and MCF-7 cell lines andevaluation of tumor suppressor gene methylation

YUNUS KASIM TERZİ, ÖZGE ÖZER KAYA, ÖZLEM DARCANSOY İŞERİ, ZERRİN ÇELİK, FERİDE İFFET ŞAHİN

A small intron of Drosophila gamma carboxylase, not predictable in silico for mammalian splicing machinery, is spliced improperly in human cells

SOLMAZ MONIRI JAVADHESARI, ALIREZA ZOMORODIPOUR, SPIDEH VALIMEHR

Eukaryotic expression, purification, identification, and tissuedistribution of porcine PID1

XIAOLING CHEN, HUAN WANG, BO ZHOU, ZHIQING HUANG, GANG JIA, GUANGMANG LIU, HUA ZHAO

Analysis of structure and differential expression of Pseudomonas syringae 5-like(RPS5-like) genes in pathogen-infected Vitis flexuosa

MD. ZAHERUL ISLAM, SOON YOUNG AHN, HAE KEUN YUN