Sulforaphane Modulates Cell Migration and Expression of β-Catenin and Epithelial Mesenchymal Transition Markers in Breast Cancer Cells
Abstract
Background: We aimed to assess the effect of sulforaphane (SFN) on breast cancer cell migration and also its effect on the expression of epithelial mesenchymal transition (EMT) markers and β-catenin.
Methods: This study was performed in Shahroud University of Medical Sciences, Shahroud, Iran from 2017- 2018. In this experimental study, MDA-MB-231 cells were treated with different concentrations of SFN (5, 10, 20, 30 and 40 μM) at different time points of 24, 48, and 72 h. The control group was untreated cells. The inhibitory effects of different concentrations of SFN on cell migration at different time points were evaluated using scratch assay. Moreover, apoptosis was assessed by using flow cytometric analysis. The expression of βcatenin and EMT markers of ZEB1, fibronectin, and claudin-1 were determined by real-time PCR. Western blotting analysis of β-catenin was applied to determine its changes after SFN treatment
Results: SFN markedly inhibited the migration of cells at concentrations of 10, 20, 30, and 40µM after 24, 48, and 72 h. At relatively, high concentrations (30, 40µM), SFN induced apoptosis. Moreover, SFN reduced the gene expression of ZEB1, fibronectin, and claudin-1 after 72 h. The expression of β-catenin revealed a time-dependent decrease at the concentration of 40 µM SFN.
Conclusion: Downregulation of EMT markers and β-catenin showed accordance with the inhibition of migration. SFN could be a promising drug candidate to reduce metastasis in breast cancer.
1. Tevaarwerk AJ, Gray RJ, Schneider BP et al (2013). Survival in patients with metastatic recurrent breast cancer after adjuvant chemotherapy: little evidence of improvement over the past 30 years. Cancer,119:1140-8.
2. Lamouille S, Xu J, Derynck R (2014). Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol,15:178-96.
3. Zhou B, Moodie A, Blanchard AA et al (2015). Claudin 1 in Breast Cancer: New Insights. J Clin Med,4:1960-76.
4. Ghahhari NM, Babashah S (2015). Interplay between microRNAs and WNT/β-catenin signalling pathway regulates epithelial–mesenchymal transition in cancer. Eur J Cancer,51:1638-49.
5. Ahmadiankia N, Moghaddam HK, Mishan MA et al (2016). Berberine suppresses migration of MCF-7 breast cancer cells through down-regulation of chemokine receptors. Iran J Basic Med Sci,19:125-31.
6. Hecht SS (2000). Inhibition of carcinogenesis by isothiocyanates. Drug Metab Rev,32:395-411.
7. Qazi A, Pal J, Maitah M et al (2010). Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy. Transl Oncol,3:389-99.
8. Mishan MA, Ahmadiankia N, Matin MM et al (2015). Role of Berberine on molecular markers involved in migration of esophageal cancer cells. Cell Mol Biol (Noisy-le-grand),61:37-43.
9. Pawlik A, Wiczk A, Kaczynska A et al (2013). Sulforaphane inhibits growth of phenotypically different breast cancer cells. Eur J Nutr,52:1949-58.
10. Mishan MA, Heirani-Tabasi A, Mokhberian N et al (2015). Analysis of Chemokine Receptor Gene Expression in Esophageal Cancer Cells Compared with Breast Cancer with Insights into Metastasis. Iran J Public Health,44:1353-8.
11. Bagheri M, Fazli M, Saeednia S et al (2018). Pomegranate peel extract inhibits expression of β-catenin, epithelial mesenchymal transition, and metastasis in triple negative breast cancer cells. Cell Mol Biol (Noisy-le-grand),64:86-91.
12. Li W, Khor TO, Xu C et al (2008). Activation of Nrf2-antioxidant signaling attenuates NFκB-inflammatory response and elicits apoptosis. Biochem Pharmacol,76:1485-9.
13. Kensler TW, Egner PA, Agyeman AS et al (2013). Keap1-nrf2 signaling: a target for cancer prevention by sulforaphane. Top Curr Chem,329:163-77.
14. Wang L, Tian Z, Yang Q et al (2015). Sulforaphane inhibits thyroid cancer cell growth and invasiveness through the reactive oxygen species-dependent pathway. Oncotarget,6:25917-31.
15. Spaderna S, Schmalhofer O, Wahlbuhl M et al (2008). The transcriptional repressor ZEB1 promotes metastasis and loss of cell polarity in cancer. Cancer Res,68:537-44.
16. Thomson S, Buck E, Petti F et al (2005). Epithelial to mesenchymal transition is a determinant of sensitivity of non–small-cell lung carcinoma cell lines and xenografts to epidermal growth factor receptor inhibition. Cancer Res,65:9455-62.
17. Sanchez-Tillo E, Fanlo L, Siles L et al (2014). The EMT activator ZEB1 promotes tumor growth and determines differential response to chemotherapy in mantle cell lymphoma. Cell Death Differ,21:247-57.
18. Fernandez-Garcia B, Eiro N, Marin L et al (2014). Expression and prognostic significance of fibronectin and matrix metalloproteases in breast cancer metastasis. Histopathology,64:512-22.
19. Konac E, Kiliccioglu I, Sogutdelen E et al (2017). Do the expressions of epithelial-mesenchymal transition proteins, periostin, integrin-alpha4 and fibronectin correlate with clinico-pathological features and prognosis of metastatic castration-resistant prostate cancer? Exp Biol Med (Maywood),242:1795-801.
20. Yi W, Xiao E, Ding R et al (2016). High expression of fibronectin is associated with poor prognosis, cell proliferation and malignancy via the NF-kappaB/p53-apoptosis signaling pathway in colorectal cancer. Oncol Rep,36:3145-53.
21. Escudero-Esparza A, Jiang WG, Martin TA (2011). The Claudin family and its role in cancer and metastasis. Front Biosci (Landmark Ed),16:1069-83.
22. Higashi Y, Suzuki S, Sakaguchi T et al (2007). Loss of claudin-1 expression correlates with malignancy of hepatocellular carcinoma. J Surg Res,139:68-76.
23. Morohashi S, Kusumi T, Sato F et al (2007). Decreased expression of claudin-1 correlates with recurrence status in breast cancer. Int J Mol Med,20:139-43.
24. Chao YC, Pan SH, Yang SC et al (2009). Claudin-1 is a metastasis suppressor and correlates with clinical outcome in lung adenocarcinoma. Am J Respir Crit Care Med,179:123-33.
25. Oku N, Sasabe E, Ueta E et al (2006). Tight junction protein claudin-1 enhances the invasive activity of oral squamous cell carcinoma cells by promoting cleavage of laminin-5 gamma2 chain via matrix metalloproteinase (MMP)-2 and membrane-type MMP-1. Cancer Res,66:5251-7.
26. Leotlela PD, Wade MS, Duray PH et al (2007). Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility. Oncogene,26:3846-56.
27. Dhawan P, Singh AB, Deane NG et al (2005). Claudin-1 regulates cellular transformation and metastatic behavior in colon cancer. J Clin Invest,115:1765-76.
28. Kinugasa T, Huo Q, Higashi D et al (2007). Selective up-regulation of claudin-1 and claudin-2 in colorectal cancer. Anticancer Res,27:3729-34.
29. Suh Y, Yoon CH, Kim Rk et al (2017). Claudin-1 induces epithelial-mesenchymal transition through activation of the c-Abl-ERK signaling pathway in human liver cells. Oncogene, 32:4873-82.
30. Chavez KJ, Garimella SV, Lipkowitz S (2010). Triple negative breast cancer cell lines: one tool in the search for better treatment of triple negative breast cancer. Breast Dis,32:35-48.
31. Kao J, Salari K, Bocanegra M et al (2009). Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PLoS One,4:e6146.
32. Blanchard AA, Ma X, Dueck KJ et al (2013). Claudin 1 expression in basal-like breast cancer is related to patient age. BMC Cancer,13:268.
33. Achari C, Winslow S, Larsson C (2015). Down Regulation of CLDND1 Induces Apoptosis in Breast Cancer Cells. PLoS One,10:e0130300.
34. Cai J, Guan H, Fang L et al (2013). MicroRNA-374a activates Wnt/beta-catenin signaling to promote breast cancer metastasis. J Clin Invest,123:566-79.
35. Orford K, Crockett C, Jensen JP et al (1997). Serine phosphorylation-regulated ubiquitina-tion and degradation of β-catenin. J Biol Chem ,272:24735-8.
36. Valenta T, Hausmann G, Basler K (2012). The many faces and functions of beta-catenin. Embo j,31:2714-36.
37. Nusse R, Clevers H (2017). Wnt/beta-Catenin Signaling, Disease, and Emerging Therapeutic Modalities. Cell,169:985-99.
38. Prosperi JR, Goss KH (2010). A Wnt-ow of opportunity: targeting the Wnt/beta-catenin pathway in breast cancer. Curr Drug Targets,11:1074-88.
39. Takahashi-Yanaga F, Kahn M (2010). Targeting Wnt signaling: can we safely eradicate cancer stem cells? Clin Cancer Res,16:3153-62.
2. Lamouille S, Xu J, Derynck R (2014). Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol,15:178-96.
3. Zhou B, Moodie A, Blanchard AA et al (2015). Claudin 1 in Breast Cancer: New Insights. J Clin Med,4:1960-76.
4. Ghahhari NM, Babashah S (2015). Interplay between microRNAs and WNT/β-catenin signalling pathway regulates epithelial–mesenchymal transition in cancer. Eur J Cancer,51:1638-49.
5. Ahmadiankia N, Moghaddam HK, Mishan MA et al (2016). Berberine suppresses migration of MCF-7 breast cancer cells through down-regulation of chemokine receptors. Iran J Basic Med Sci,19:125-31.
6. Hecht SS (2000). Inhibition of carcinogenesis by isothiocyanates. Drug Metab Rev,32:395-411.
7. Qazi A, Pal J, Maitah M et al (2010). Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy. Transl Oncol,3:389-99.
8. Mishan MA, Ahmadiankia N, Matin MM et al (2015). Role of Berberine on molecular markers involved in migration of esophageal cancer cells. Cell Mol Biol (Noisy-le-grand),61:37-43.
9. Pawlik A, Wiczk A, Kaczynska A et al (2013). Sulforaphane inhibits growth of phenotypically different breast cancer cells. Eur J Nutr,52:1949-58.
10. Mishan MA, Heirani-Tabasi A, Mokhberian N et al (2015). Analysis of Chemokine Receptor Gene Expression in Esophageal Cancer Cells Compared with Breast Cancer with Insights into Metastasis. Iran J Public Health,44:1353-8.
11. Bagheri M, Fazli M, Saeednia S et al (2018). Pomegranate peel extract inhibits expression of β-catenin, epithelial mesenchymal transition, and metastasis in triple negative breast cancer cells. Cell Mol Biol (Noisy-le-grand),64:86-91.
12. Li W, Khor TO, Xu C et al (2008). Activation of Nrf2-antioxidant signaling attenuates NFκB-inflammatory response and elicits apoptosis. Biochem Pharmacol,76:1485-9.
13. Kensler TW, Egner PA, Agyeman AS et al (2013). Keap1-nrf2 signaling: a target for cancer prevention by sulforaphane. Top Curr Chem,329:163-77.
14. Wang L, Tian Z, Yang Q et al (2015). Sulforaphane inhibits thyroid cancer cell growth and invasiveness through the reactive oxygen species-dependent pathway. Oncotarget,6:25917-31.
15. Spaderna S, Schmalhofer O, Wahlbuhl M et al (2008). The transcriptional repressor ZEB1 promotes metastasis and loss of cell polarity in cancer. Cancer Res,68:537-44.
16. Thomson S, Buck E, Petti F et al (2005). Epithelial to mesenchymal transition is a determinant of sensitivity of non–small-cell lung carcinoma cell lines and xenografts to epidermal growth factor receptor inhibition. Cancer Res,65:9455-62.
17. Sanchez-Tillo E, Fanlo L, Siles L et al (2014). The EMT activator ZEB1 promotes tumor growth and determines differential response to chemotherapy in mantle cell lymphoma. Cell Death Differ,21:247-57.
18. Fernandez-Garcia B, Eiro N, Marin L et al (2014). Expression and prognostic significance of fibronectin and matrix metalloproteases in breast cancer metastasis. Histopathology,64:512-22.
19. Konac E, Kiliccioglu I, Sogutdelen E et al (2017). Do the expressions of epithelial-mesenchymal transition proteins, periostin, integrin-alpha4 and fibronectin correlate with clinico-pathological features and prognosis of metastatic castration-resistant prostate cancer? Exp Biol Med (Maywood),242:1795-801.
20. Yi W, Xiao E, Ding R et al (2016). High expression of fibronectin is associated with poor prognosis, cell proliferation and malignancy via the NF-kappaB/p53-apoptosis signaling pathway in colorectal cancer. Oncol Rep,36:3145-53.
21. Escudero-Esparza A, Jiang WG, Martin TA (2011). The Claudin family and its role in cancer and metastasis. Front Biosci (Landmark Ed),16:1069-83.
22. Higashi Y, Suzuki S, Sakaguchi T et al (2007). Loss of claudin-1 expression correlates with malignancy of hepatocellular carcinoma. J Surg Res,139:68-76.
23. Morohashi S, Kusumi T, Sato F et al (2007). Decreased expression of claudin-1 correlates with recurrence status in breast cancer. Int J Mol Med,20:139-43.
24. Chao YC, Pan SH, Yang SC et al (2009). Claudin-1 is a metastasis suppressor and correlates with clinical outcome in lung adenocarcinoma. Am J Respir Crit Care Med,179:123-33.
25. Oku N, Sasabe E, Ueta E et al (2006). Tight junction protein claudin-1 enhances the invasive activity of oral squamous cell carcinoma cells by promoting cleavage of laminin-5 gamma2 chain via matrix metalloproteinase (MMP)-2 and membrane-type MMP-1. Cancer Res,66:5251-7.
26. Leotlela PD, Wade MS, Duray PH et al (2007). Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility. Oncogene,26:3846-56.
27. Dhawan P, Singh AB, Deane NG et al (2005). Claudin-1 regulates cellular transformation and metastatic behavior in colon cancer. J Clin Invest,115:1765-76.
28. Kinugasa T, Huo Q, Higashi D et al (2007). Selective up-regulation of claudin-1 and claudin-2 in colorectal cancer. Anticancer Res,27:3729-34.
29. Suh Y, Yoon CH, Kim Rk et al (2017). Claudin-1 induces epithelial-mesenchymal transition through activation of the c-Abl-ERK signaling pathway in human liver cells. Oncogene, 32:4873-82.
30. Chavez KJ, Garimella SV, Lipkowitz S (2010). Triple negative breast cancer cell lines: one tool in the search for better treatment of triple negative breast cancer. Breast Dis,32:35-48.
31. Kao J, Salari K, Bocanegra M et al (2009). Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PLoS One,4:e6146.
32. Blanchard AA, Ma X, Dueck KJ et al (2013). Claudin 1 expression in basal-like breast cancer is related to patient age. BMC Cancer,13:268.
33. Achari C, Winslow S, Larsson C (2015). Down Regulation of CLDND1 Induces Apoptosis in Breast Cancer Cells. PLoS One,10:e0130300.
34. Cai J, Guan H, Fang L et al (2013). MicroRNA-374a activates Wnt/beta-catenin signaling to promote breast cancer metastasis. J Clin Invest,123:566-79.
35. Orford K, Crockett C, Jensen JP et al (1997). Serine phosphorylation-regulated ubiquitina-tion and degradation of β-catenin. J Biol Chem ,272:24735-8.
36. Valenta T, Hausmann G, Basler K (2012). The many faces and functions of beta-catenin. Embo j,31:2714-36.
37. Nusse R, Clevers H (2017). Wnt/beta-Catenin Signaling, Disease, and Emerging Therapeutic Modalities. Cell,169:985-99.
38. Prosperi JR, Goss KH (2010). A Wnt-ow of opportunity: targeting the Wnt/beta-catenin pathway in breast cancer. Curr Drug Targets,11:1074-88.
39. Takahashi-Yanaga F, Kahn M (2010). Targeting Wnt signaling: can we safely eradicate cancer stem cells? Clin Cancer Res,16:3153-62.
| Files | ||
| Issue | Vol 49 No 1 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v49i1.3054 | |
| Keywords | ||
| Sulforaphane; Metastasis; Breast cancer EMT; β-catenin | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
BAGHERI M, FAZLI M, SAEEDNIA S, GHOLAMI KHARANAGH M, AHMADIANKIA N. Sulforaphane Modulates Cell Migration and Expression of β-Catenin and Epithelial Mesenchymal Transition Markers in Breast Cancer Cells. Iran J Public Health. 2020;49(1):77-85.
