Silibinin upregulates E-cadherin expression in MKN-45 human gastric cancer cells

Document Type: Original paper

Authors

1 Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran. Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

3 Colorectal Research Center, Iran University of Medical Sciences, Tehran, Iran.

Abstract

Background and objectives:  Gastric cancer is currently known as one of the most important causes of cancer-driven death all over the world. In patients with gastric cancer, a significant proportion of death occurs due to metastasis. On the other hand, down modulated E-cadherin level has been reported as an important contributor to tumor cell invasion and metastasis. In this regard, the present work was aimed to evaluate the impact of silibinin, a flavonolignan with established anti-tumor efficacy, on cell viability and E-cadherin expression in the gastric cancer cell line MKN-45. Methods: To determine cell viability, MTT assay was performed 48 h after silibinin treatment (at concentrations of 100, 200 and 400 μM). In addition, quantitative real-time PCR was done following total RNA extraction and cDNA synthesis, to assess E-cadherin level in cells treated with silibinin. Results: The MTT results showed concentration-dependent reducing effect of silibinin on viability of MKN-45 cells. The findings of quantitative real-time PCR analysis demonstrated upregulated E-cadherin expression in cells treated with silibinin (significantly (p≤ 0.05) at concentration of 200 μM) compared to the control cells. Conclusions: The current study suggested that silibinin may exert anti-migratory/invasive effects on gastric cancer cells by enhancing E-cadherin expression, which needs to be further investigated.
 

Keywords


[1] Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002; 2(6): 442-454.

[2] Gumbiner BM. Regulation of cadherin-mediated adhesion in morphogenesis. Nature reviews. Mol Cell Biol. 2005; 6(8): 622-634.

[3] Huiping C, Kristjansdottir S, Jonasson JG, Magnusson J, Egilsson V, Ingvarsson S. Alterations of E-cadherin and β-catenin in gastric cancer. BMC Cancer. 2001; 1(1): 1-16.

[4] Liu YN, Lee WW, Wang CY, Chao TH, Chen Y, Chen JH. Regulatory mechanisms controlling human E-cadherin gene expression. Oncogene. 2005; 24(56): 8277-8290.

[5] Oloumi A, McPhee T, Dedhar S. Regulation of E-cadherin expression and β-catenin/Tcf transcriptional activity by the integrin-linked kinase. BBA Mol Cell Res. 2004; 1691(1): 1-15.

[6] Kwon O, Jeong SJ, Kim SO, He L, Lee HG, Jang KL, Osada H, Jung M, Kim BY, Ahn JS. Modulation of E-cadherin expression by K-Ras; involvement of DNA methyltransferase-3b. Carcinogenesis. 2010; 31(7): 1194-1201.

[7] Aghdassi A, Sendler M, Guenther A, Mayerle J, Behn CO, Heidecke CD, Friess H, Büchler M, Evert M, Lerch MM, Weiss FU. Recruitment of histone deacetylases HDAC1 and HDAC2 by the transcriptional repressor ZEB1 downregulates E-cadherin expression in pancreatic cancer. Gut. 2012; 61(3): 439-448.

[8] Espada J, Peinado H, Lopez-Serra L, Setien F, Lopez-Serra P, Portela A, Renart J, Carrasco E, Calvo M, Juarranz A, Cano A. Regulation of SNAIL1 and E-cadherin function by DNMT1 in a DNA methylation-independent context. Nucleic Acids Res. 2011; 39(21): 9194-9205.

[9] Liao W, Jordaan G, Srivastava MK, Dubinett S, Sharma S, Sharma S. Effect of epigenetic histone modifications on E-cadherin splicing and expression in lung cancer. Am J Cancer Res. 2013; 3(4): 374-389.

[10] Marks PA, Rifkind RA, Richon VM, Breslow R, Miller T, Kelly WK. Histone deacetylases and cancer: causes and therapies. Nat Rev Cancer. 2001; 1(3): 194-202.

[11] Jin B, Robertson KD. DNA methyltransferases, DNA damage repair, and cancer. In: Karpf A, Ed. Epigenetic alterations in oncogenesis. advances in experimental medicine and biology. New York: Springer, 2013.

[12] Faghihloo E, Akbari A, Adjaminezhad-Fard F, Mokhtari-Azad T. Transcriptional regulation of E-cadherin and oncoprotein E7 by valproic acid in HPV positive cell lines. Iran J Basic Med Sci. 2016; 19(6): 601-607.

[13] Takai N, Narahara H. Human endometrial and ovarian cancer cells: histone deacetylase inhibitors exhibit antiproliferative activity, potently induce cell cycle arrest, and stimulate apoptosis. Curr Med Chem. 2007; 14(24): 2548-2553.

[14] Anestopoulos I, Sfakianos AP, Franco R, Chlichlia K, Panayiotidis MI, Kroll DJ, Pappa A. A novel role of silibinin as a putative epigenetic modulator in human prostate carcinoma. Molecules. 2016; Article ID 28042859.

[15] Agarwal C, Wadhwa R, Deep G, Biedermann D, Gažák R, Křen V, Agarwal R. Anti-cancer efficacy of silybin derivatives-a structure-activity relationship. PLoS One. 2013; Article ID 23555889.

[16] Kalemci S, Topal Y, Celik SY, Yilmaz N, Beydilli H, Kosar MI, Dirican N, Altuntas I. Silibinin attenuates methotrexate-induced pulmonary injury by targeting oxidative stress. Exp Ther Med. 2015; 10(2): 503-507.

[17] Jiang C, Jin S, Jiang Z, Wang J. Inhibitory effects of silibinin on proliferation and lung metastasis of human high metastasis cell line of salivary gland adenoid cystic carcinoma via autophagy induction. Oncotargets Ther. 2016; 9: 6609-6618.

[18] Bremnes RM, Veve R, Hirsch FR, Franklin WA. The E-cadherin cell-cell adhesion complex and lung cancer invasion, metastasis, and prognosis. Lung Cancer. 2002; 36(2): 115-124.

[19] Ozawa M, Kobayashi W. Cadherin cytoplasmic domains inhibit the cell surface localization of endogenous E-cadherin, blocking desmosome and tight junction formation and inducing cell dissociation. PloS One. 2014; Article ID 25121615.

[20] Van Roy F, Berx G. The cell-cell adhesion molecule E-cadherin. Cell Mol Life Sci. 2008; 65(23): 3756-3788.

[21] Petrova YI, Schecterson L, Gumbiner BM. Roles for E-cadherin cell surface regulation in cancer. Mol Biol Cell. 2016; 27(21): 3233-3244.

[22] Rugge M, Fassan M, Graham D. Epidemiology of gastric cancer. In: Strong V, Ed. Gastric cancer. Cham: Springer, 2015.

[23] Faghihloo E, Saremi MR, Mahabadi M, Akbari H,  Saberfar E. Prevalence and characteristics of Epstein-Barr virus-associated gastric cancer in Iran. Arch Iran Med. 2014; 17(11): 767-770.

[24] Yonemura Y, Bandou E, Kinoshita K, Kawamura T, Takahashi S, Endou Y, Sasaki T. Effective therapy for peritoneal dissemination in gastric cancer. Surg Oncol Clin N Am. 2003; 12(3): 635-648.

[25] Thomassen I, Gestel YR, Ramshorst B, Luyer MD, Bosscha K, Nienhuijs SW, Lemmens VE, Hingh IH. Peritoneal carcinomatosis of gastric origin: a population‐based study on incidence, survival and risk factors. Int J Cancer. 2014; 134(3): 622-628.

[26] Deep G, Gangar SC, Agarwal C, Agarwal R. Role of E-cadherin in antimigratory and antiinvasive efficacy of silibinin in prostate cancer cells. Cancer Prev Res. 2011; 4(8): 1222-1232.

[27] Mateen S, Raina K, Agarwal C, Chan D, Agarwal R. Silibinin synergizes with histone deacetylase and DNA methyltransferase inhibitors in upregulating E-cadherin expression together with inhibition of migration and invasion of human non-small cell lung cancer cells. J Pharmacol Exp Ther. 2013; 345(2): 206-214.

[28] Subramaniam D, Thombre R, Dhar A, Anant S. DNA methyltransferases: a novel target for prevention and therapy. Front Oncol. 2014; 4:80.