Does broccoli (brassica oleracea var. italica) extract have a potential therapeutic targets on human breast cancer (MDA-MB-231) using AgNOR detection method?

Gözde Çobanoğlu; Mustafa Nisari; Mehtap Nisari; Sümeyye Uçar; Gülderen Kerek; Neriman İnanç

doi:10.7197/cmj.1297877

Research Article

Year 2023, Volume: 45 Issue: 3, 44 - 56, 30.09.2023

Gözde Çobanoğlu , Mustafa Nisari , Mehtap Nisari , Sümeyye Uçar , Gülderen Kerek , Neriman İnanç

https://doi.org/10.7197/cmj.1297877

Abstract

Supporting Institution

Nuh Naci Yazgan University

Project Number

2021/SA-LTP-3

References

1. Ozmen V, Boylu S, Ok E, et al. Factors affecting breast cancer treatment delay in Turkey: a study from Turkish Federation of Breast Diseases Societies. The European Journal of Public Health. 2015;25(1):9-14.
2. Jiang X, Jiang X, Liu Y, et al. Chemopreventive activity of sulforaphane. Drug design, development and therapy. 2018: 2905-13.
3. Cheung KL, Kong A-N. Molecular targets of dietary phenethyl isothiocyanate and sulforaphane for cancer chemoprevention. The AAPS journal. 2010; 12:87-97.
4. Akhlaghi M, Bandy B. Dietary broccoli sprouts protect against myocardial oxidative damage and cell death during ischemia-reperfusion. Plant Foods for Human Nutrition. 2010; 65:193-9.
5. Moon J-K, Kim J-R, Ahn Y-J, Shibamoto T. Analysis and anti-Helicobacter activity of sulforaphane and related compounds present in broccoli (Brassica oleracea L.) sprouts. Journal of Agricultural and Food Chemistry. 2010;58(11):6672-7.
6. Baenas N, Gómez-Jodar I, Moreno DA, García-Viguera C, Periago PM. Broccoli and radish sprouts are safe and rich in bioactive phytochemicals. Postharvest Biology and Technology. 2017; 127:60-7.
7. Le TN, Chiu C-H, Hsieh P-C. Bioactive compounds and bioactivities of Brassica oleracea L. var. italica sprouts and microgreens: An updated overview from a nutraceutical perspective. Plants. 2020;9(8):946.
8. Roussel P, Hernandez-Verdun D. Identification of Ag-NOR proteins, markers of proliferation related to ribosomal gene activity. Experimental cell research. 1994;214(2):465-72.
9. Nisari M, Eroz R, Ertekin T, et al. Investigation of argyrophilic nucleolar organizing region. BRATISLAVA MEDICAL JOURNAL-BRATISLAVSKE LEKARSKE LISTY. 2016;117(6).
10. Goodpasture C, Bloom SE. Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma. 1975;53(1):37-50.
11. Trere D. AgNOR staining and quantification. Micron. 2000;31(2):127-31.
12. Lindner LE. Improvements in the silver-staining technique for nucleolar organizer regions (AgNOR). Journal of Histochemistry & Cytochemistry. 1993;41(3):439-45.
13. Baykara O. Current Approaches in Cancer Treatment. Balikesir Journal of Health Sciences. 2016; 5(3): 154-165.
14. Ullah MF, Bhat SH, Husain E, et al. Cancer chemopreventive pharmacology of phytochemicals derived from plants of dietary and non-dietary origin: implication for alternative and complementary approaches. Phytochemistry Reviews. 2014; 13:811-33.
15. Abuajah CI, Ogbonna AC, Osuji CM. Functional components and medicinal properties of food: a review. Journal of food science and technology. 2015; 52:2522-9 . 16. Ambrosone CB, McCann SE, Freudenheim JL, et al. Breast cancer risk in premenopausal women is inversely associated with consumption of broccoli, a source of isothiocyanates, but is not modified by GST genotype. The Journal of nutrition. 2004;134(5):1134-8.
17. Fimognari C, Hrelia P. Sulforaphane as a promising molecule for fighting cancer. Mutation Research/Reviews in Mutation Research. 2007;635(2-3):90-104.
18. López-Chillón MT, Carazo-Díaz C, Prieto-Merino D, et al. Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects. Clinical Nutrition. 2019;38(2):745-52.
19. Mao F, Xiao B, Jiang Z, et al. J. Anticancer effect of Lycium barbarum polysaccharides on colon cancer cells involves G0/G1 phase arrest. Medical Oncology. 2011; 28:121-6.
20. Pham N-A, Jacobberger JW, Schimmer AD, et al. The dietary isothiocyanate sulforaphane targets pathways of apoptosis, cell cycle arrest, and oxidative stress in human pancreatic cancer cells and inhibits tumor growth in severe combined immunodeficient mice. Molecular cancer therapeutics. 2004;3(10):1239-48.
21. Cornblatt BS, Ye L, Dinkova-Kostova AT, et al. Preclinical and clinical evaluation of sulforaphane for chemoprevention in the breast. Carcinogenesis. 2007;28(7):1485-90.
22. Kallifatidis G, Rausch V, Baumann B, et al. Sulforaphane targets pancreatic tumour-initiating cells by NF-κB-induced antiapoptotic signalling. Gut. 2009;58(7):949-63.
23. Myzak MC, Dashwood RH. Chemoprotection by sulforaphane: keep one eye beyond Keap1. Cancer letters. 2006;233(2):208-18.
24. Licznerska B, Szaefer H, Matuszak I, Murias M, and Baer-Dubowska W. Modulating Potential of L‐Sulforaphane in the Expression of Cytochrome P450 to Identify Potential Targets for Breast Cancer Chemoprevention and Therapy Using Breast Cell Lines. Phytotherapy Research. 2015;29(1):93-9.
25. Lewinska A, Adamczyk-Grochala J, Deregowska A, and Wnuk M. Sulforaphane-induced cell cycle arrest and senescence are accompanied by DNA hypomethylation and changes in microRNA profile in breast cancer cells. Theranostics. 2017;7(14):3461.
26. Licznerska B, Szaefer H, Krajka-Kuźniak V. R-sulforaphane modulates the expression profile of AhR, ERα, Nrf2, NQO1, and GSTP in human breast cell lines. Molecular and Cellular Biochemistry. 2021; 476:525-33.
27. Pledgie-Tracy A, Sobolewski MD, Davidson NE. Sulforaphane induces cell type–specific apoptosis in human breast cancer cell lines. Molecular cancer therapeutics. 2007;6(3):1013-21.
28. Paśko P, Tyszka-Czochara M, Galanty A, et al. Comparative study of predominant phytochemical compounds and proapoptotic potential of broccoli sprouts and florets. Plant Foods for Human Nutrition. 2018; 73:95-100.
29. Kim H-J, You M-K, Lee Y-H, et al. Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells. Nutrition research and practice. 2018;12(6):494-502.
30. Lin HW, Hsieh MJ, Yeh CB, et al. Coronarin D induces apoptotic cell death through the JNK pathway in human hepatocellular carcinoma. Environmental toxicology. 2018;33(9):946-54.
31. Jackson SJ, Singletary KW. Sulforaphane: a naturally occurring mammary carcinoma mitotic inhibitor, which disrupts tubulin polymerization. Carcinogenesis. 2004;25(2):219-27.
32. Cheng A-C, Shen C-J, Hung C-M, et al. C. Sulforaphane decrease of SERTAD1 expression triggers G1/S arrest in breast cancer cells. Journal of medicinal food. 2019;22(5):444-50.
33. Rutz J, Juengel E, Euler S, et al. Chronic sulforaphane application does not induce resistance in renal cell carcinoma cells. Anticancer Research. 2018;38(11):6201-7.
34. Liu K-C, Shih T-Y, Kuo C-L, et al. Sulforaphane induces cell death through G2/M phase arrest and triggers apoptosis in HCT 116 human colon cancer cells. The American Journal of Chinese Medicine. 2016;44(06):1289-310.
35. Mao F, Xiao B, Jiang Z, Zhao J, Huang X, Guo J. Anticancer effect of Lycium barbarum polysaccharides on colon cancer cells involves G0/G1 phase arrest. Medical Oncology. 2011; 28:121-6.
36. Ertekin T, Bozkurt O, Eroz R, Nisari M, Bircan D, Nisari M, Unur E. May argyrophilic nucleolar organizing region–associated protein synthesis be used for selection of the most reliable dose of the drugs such as Rhamnetin in cancer treatments. Bratislava Medical Journal. 2016;117(11):653–658.
37. Nisari M, Eröz R. Does capsaicin have therapeutic benefits in human colon adenocarcinoma? Selection of the most reliable dose via AgNOR. Turkish Journal of Medical Sciences 2020; 50: 1076-1081.

Does broccoli (brassica oleracea var. italica) extract have a potential therapeutic targets on human breast cancer (MDA-MB-231) using AgNOR detection method?

Year 2023, Volume: 45 Issue: 3, 44 - 56, 30.09.2023

Gözde Çobanoğlu , Mustafa Nisari , Mehtap Nisari , Sümeyye Uçar , Gülderen Kerek , Neriman İnanç

https://doi.org/10.7197/cmj.1297877

Abstract

In this study, it was aimed to determine the in vitro effect of the extract obtained from mature broccoli on the breast cancer cell line. MDA-MB231 breast cancer cell line was used in the study. MDA-MB-231 cells were exposed to broccoli extract at 37°C and 5% CO2 for varying durations (24 and 48 hours) and doses (125 and 250 µl/ml).At the end of the incubation period, viability, apoptosis, cell cycle and AgNOR protein status of MDA-MB-231 cells were examined in the Muse Cell Analyzer. In the groups containing broccoli extract, a decrease in the percentage of viable cells and a significant increase in the percentage of early and total apoptosis were observed for both doses compared to the control. In the cell cycle test, the number of cells in the S phase increased in all groups.It was observed that the groups containing broccoli extract slowed down the cell cycle in the transition to the S checkpoint. AgNOR staining results also supported cell cycle and apopitosis, and AgNOR number and TAA/NA ratio decreased in the 125 µl/ml broccoli extract group after 24-48 hours and were found to be statistically significant compared to the control group. It was determined that broccoli increased apoptosis on breast cancer cells by various mechanisms and inhibited cell viability/cell growth. The results were similar to the results of AgNOR protein synthesis. The study showed that the regular and correct consumption of broccoli could be effective in preventing cancer formation and slowing its progression.

Keywords

Broccoli, MDA-MB-231, AgNOR staining

Project Number

2021/SA-LTP-3

References

1. Ozmen V, Boylu S, Ok E, et al. Factors affecting breast cancer treatment delay in Turkey: a study from Turkish Federation of Breast Diseases Societies. The European Journal of Public Health. 2015;25(1):9-14.
2. Jiang X, Jiang X, Liu Y, et al. Chemopreventive activity of sulforaphane. Drug design, development and therapy. 2018: 2905-13.
3. Cheung KL, Kong A-N. Molecular targets of dietary phenethyl isothiocyanate and sulforaphane for cancer chemoprevention. The AAPS journal. 2010; 12:87-97.
4. Akhlaghi M, Bandy B. Dietary broccoli sprouts protect against myocardial oxidative damage and cell death during ischemia-reperfusion. Plant Foods for Human Nutrition. 2010; 65:193-9.
5. Moon J-K, Kim J-R, Ahn Y-J, Shibamoto T. Analysis and anti-Helicobacter activity of sulforaphane and related compounds present in broccoli (Brassica oleracea L.) sprouts. Journal of Agricultural and Food Chemistry. 2010;58(11):6672-7.
6. Baenas N, Gómez-Jodar I, Moreno DA, García-Viguera C, Periago PM. Broccoli and radish sprouts are safe and rich in bioactive phytochemicals. Postharvest Biology and Technology. 2017; 127:60-7.
7. Le TN, Chiu C-H, Hsieh P-C. Bioactive compounds and bioactivities of Brassica oleracea L. var. italica sprouts and microgreens: An updated overview from a nutraceutical perspective. Plants. 2020;9(8):946.
8. Roussel P, Hernandez-Verdun D. Identification of Ag-NOR proteins, markers of proliferation related to ribosomal gene activity. Experimental cell research. 1994;214(2):465-72.
9. Nisari M, Eroz R, Ertekin T, et al. Investigation of argyrophilic nucleolar organizing region. BRATISLAVA MEDICAL JOURNAL-BRATISLAVSKE LEKARSKE LISTY. 2016;117(6).
10. Goodpasture C, Bloom SE. Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma. 1975;53(1):37-50.
11. Trere D. AgNOR staining and quantification. Micron. 2000;31(2):127-31.
12. Lindner LE. Improvements in the silver-staining technique for nucleolar organizer regions (AgNOR). Journal of Histochemistry & Cytochemistry. 1993;41(3):439-45.
13. Baykara O. Current Approaches in Cancer Treatment. Balikesir Journal of Health Sciences. 2016; 5(3): 154-165.
14. Ullah MF, Bhat SH, Husain E, et al. Cancer chemopreventive pharmacology of phytochemicals derived from plants of dietary and non-dietary origin: implication for alternative and complementary approaches. Phytochemistry Reviews. 2014; 13:811-33.
15. Abuajah CI, Ogbonna AC, Osuji CM. Functional components and medicinal properties of food: a review. Journal of food science and technology. 2015; 52:2522-9 . 16. Ambrosone CB, McCann SE, Freudenheim JL, et al. Breast cancer risk in premenopausal women is inversely associated with consumption of broccoli, a source of isothiocyanates, but is not modified by GST genotype. The Journal of nutrition. 2004;134(5):1134-8.
17. Fimognari C, Hrelia P. Sulforaphane as a promising molecule for fighting cancer. Mutation Research/Reviews in Mutation Research. 2007;635(2-3):90-104.
18. López-Chillón MT, Carazo-Díaz C, Prieto-Merino D, et al. Effects of long-term consumption of broccoli sprouts on inflammatory markers in overweight subjects. Clinical Nutrition. 2019;38(2):745-52.
19. Mao F, Xiao B, Jiang Z, et al. J. Anticancer effect of Lycium barbarum polysaccharides on colon cancer cells involves G0/G1 phase arrest. Medical Oncology. 2011; 28:121-6.
20. Pham N-A, Jacobberger JW, Schimmer AD, et al. The dietary isothiocyanate sulforaphane targets pathways of apoptosis, cell cycle arrest, and oxidative stress in human pancreatic cancer cells and inhibits tumor growth in severe combined immunodeficient mice. Molecular cancer therapeutics. 2004;3(10):1239-48.
21. Cornblatt BS, Ye L, Dinkova-Kostova AT, et al. Preclinical and clinical evaluation of sulforaphane for chemoprevention in the breast. Carcinogenesis. 2007;28(7):1485-90.
22. Kallifatidis G, Rausch V, Baumann B, et al. Sulforaphane targets pancreatic tumour-initiating cells by NF-κB-induced antiapoptotic signalling. Gut. 2009;58(7):949-63.
23. Myzak MC, Dashwood RH. Chemoprotection by sulforaphane: keep one eye beyond Keap1. Cancer letters. 2006;233(2):208-18.
24. Licznerska B, Szaefer H, Matuszak I, Murias M, and Baer-Dubowska W. Modulating Potential of L‐Sulforaphane in the Expression of Cytochrome P450 to Identify Potential Targets for Breast Cancer Chemoprevention and Therapy Using Breast Cell Lines. Phytotherapy Research. 2015;29(1):93-9.
25. Lewinska A, Adamczyk-Grochala J, Deregowska A, and Wnuk M. Sulforaphane-induced cell cycle arrest and senescence are accompanied by DNA hypomethylation and changes in microRNA profile in breast cancer cells. Theranostics. 2017;7(14):3461.
26. Licznerska B, Szaefer H, Krajka-Kuźniak V. R-sulforaphane modulates the expression profile of AhR, ERα, Nrf2, NQO1, and GSTP in human breast cell lines. Molecular and Cellular Biochemistry. 2021; 476:525-33.
27. Pledgie-Tracy A, Sobolewski MD, Davidson NE. Sulforaphane induces cell type–specific apoptosis in human breast cancer cell lines. Molecular cancer therapeutics. 2007;6(3):1013-21.
28. Paśko P, Tyszka-Czochara M, Galanty A, et al. Comparative study of predominant phytochemical compounds and proapoptotic potential of broccoli sprouts and florets. Plant Foods for Human Nutrition. 2018; 73:95-100.
29. Kim H-J, You M-K, Lee Y-H, et al. Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells. Nutrition research and practice. 2018;12(6):494-502.
30. Lin HW, Hsieh MJ, Yeh CB, et al. Coronarin D induces apoptotic cell death through the JNK pathway in human hepatocellular carcinoma. Environmental toxicology. 2018;33(9):946-54.
31. Jackson SJ, Singletary KW. Sulforaphane: a naturally occurring mammary carcinoma mitotic inhibitor, which disrupts tubulin polymerization. Carcinogenesis. 2004;25(2):219-27.
32. Cheng A-C, Shen C-J, Hung C-M, et al. C. Sulforaphane decrease of SERTAD1 expression triggers G1/S arrest in breast cancer cells. Journal of medicinal food. 2019;22(5):444-50.
33. Rutz J, Juengel E, Euler S, et al. Chronic sulforaphane application does not induce resistance in renal cell carcinoma cells. Anticancer Research. 2018;38(11):6201-7.
34. Liu K-C, Shih T-Y, Kuo C-L, et al. Sulforaphane induces cell death through G2/M phase arrest and triggers apoptosis in HCT 116 human colon cancer cells. The American Journal of Chinese Medicine. 2016;44(06):1289-310.
35. Mao F, Xiao B, Jiang Z, Zhao J, Huang X, Guo J. Anticancer effect of Lycium barbarum polysaccharides on colon cancer cells involves G0/G1 phase arrest. Medical Oncology. 2011; 28:121-6.
36. Ertekin T, Bozkurt O, Eroz R, Nisari M, Bircan D, Nisari M, Unur E. May argyrophilic nucleolar organizing region–associated protein synthesis be used for selection of the most reliable dose of the drugs such as Rhamnetin in cancer treatments. Bratislava Medical Journal. 2016;117(11):653–658.
37. Nisari M, Eröz R. Does capsaicin have therapeutic benefits in human colon adenocarcinoma? Selection of the most reliable dose via AgNOR. Turkish Journal of Medical Sciences 2020; 50: 1076-1081.

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Details

Primary Language	English
Subjects	Health Care Administration
Journal Section	Basic Science Research Articles
Authors	Gözde Çobanoğlu 0000-0002-7040-6212 Mustafa Nisari 0000-0001-7469-8921 Mehtap Nisari 0000-0002-1126-7478 Sümeyye Uçar 0000-0003-3378-3745 Gülderen Kerek 0000-0002-8582-0395 Neriman İnanç 0000-0001-5026-4133
Project Number	2021/SA-LTP-3
Publication Date	September 30, 2023
Acceptance Date	September 27, 2023
Published in Issue	Year 2023Volume: 45 Issue: 3

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AMA	Çobanoğlu G, Nisari M, Nisari M, Uçar S, Kerek G, İnanç N. Does broccoli (brassica oleracea var. italica) extract have a potential therapeutic targets on human breast cancer (MDA-MB-231) using AgNOR detection method?. CMJ. September 2023;45(3):44-56. doi:10.7197/cmj.1297877

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