Research Article
Quercetin'in HT-29 ve HCT-116 kolon kanseri hücre hatları üzerine etkisinin RIPK1, RIPK3 ve MLKL genlerinin ekspresyonları ile incelenmesi
Abstract
Amaç: Quercetin kolon kanseri dahil birçok kanser çeşidinde anti-kanser aktivite gösteren bir bileşiktir. Ancak, quercetinin nekroptoz yolağı üzerine etkilerini gösteren çalışmalar kısıtlıdır ve bu nedenle bu çalışmada quercetinin nekroptoz yolağına etkisinin belirlenmesi amaçlanmıştır.
Gereç ve Yöntem: HT-29 ve HCT-116 kolon kanseri hücreleri kültür edilip farklı konsantrasyondaki quercetinin hücre canlılığına etkisi MTT yöntemi ile belirlendi. Sonrasında quercetinin nekroptoza etkisinin belirlenmesi için RIPK1, RIPK3 ve MLKL genlerinin ekspresyon seviyesi analiz edildi.
Bulgular: HT-29 hücrelerinde quercetinin aktif dozu 50 µM (p=0,0286) olarak bulunurken HCT-116 hücrelerinde 100 µM (p=0,009) bulundu. 50 ve 100 µM quercetin ile maruz bırakılan HT-29 hücrelerinde nekroptoz belirteçlerinin ekspresyon seviyesinde ciddi bir artış tespit edildi.
Sonuç: Bu çalışmanın sonuçları quercetinin nekroptoz yolağının aktif bir düzenleyicisi olabileceğini göstermiştir.
Ethical Statement
Bu çalışma için etik kurul onayı etik kurul izni alınmamıştır.
Supporting Institution
Bu çalışma, 2209-A Üniversite Öğrencileri Araştırma Projeleri Destekleme Programı kapsamında Adıyaman Üniversitesi, Sağlık Hizmetleri Meslek Yüksekokulu, Tıbbi Laboratuvar Teknikleri Programı öğrencisi Alp Can TUNCER’in 1919B012202294 numaralı projesi ile Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK) tarafından desteklenmiştir.
Project Number
1919B012202294
References
- Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2021;71(3):209-249.
- Darband SG, Kaviani M, Yousefi B, et al. Quercetin: A functional dietary flavonoid with potential chemo-preventive properties in colorectal cancer. J Cell Physiol. Sep 2018;233(9):6544-6560. doi:10.1002/jcp.26595
- Aoyama N, Kawado M, Yamada H, et al. Low intake of vegetables and fruits and risk of colorectal cancer: the Japan Collaborative Cohort Study. Journal of epidemiology. 2014;24(5):353-360.
- Turati F, Rossi M, Pelucchi C, Levi F, La Vecchia C. Fruit and vegetables and cancer risk: a review of southern European studies. British Journal of Nutrition. 2015;113(S2):S102-S110.
- Sak K. Site-specific anticancer effects of dietary flavonoid quercetin. Nutrition cancer. 2014;66(2):177-193.
- van Erk MJ, Roepman P, van der Lende TR, et al. Integrated assessment by multiple gene expression analysis of quercetin bioactivity on anticancer–related mechanisms in colon cancer cells in vitro. European journal of nutrition. 2005;44(3):143-156.
- Kim H-J, Kim S-K, Kim B-S, et al. Apoptotic effect of quercetin on HT-29 colon cancer cells via the AMPK signaling pathway. Journal of agricultural food chemistry. 2010;58(15):8643-8650.
- Wenzel U, Herzog A, Kuntz S, Daniel H. Protein expression profiling identifies molecular targets of quercetin as a major dietary flavonoid in human colon cancer cells. Proteomics. 2004;4(7):2160-2174.
- Kim H-S, Wannatung T, Lee S, et al. Quercetin enhances hypoxia-mediated apoptosis via direct inhibition of AMPK activity in HCT116 colon cancer. Apoptosis. 2012;17(9):938-949.
- Zhang H, Zhang M, Yu L, Zhao Y, He N, Yang X. Antitumor activities of quercetin and quercetin-5′, 8-disulfonate in human colon and breast cancer cell lines. Food Chemical Toxicology. 2012;50(5):1589-1599.
- He S, Wang L, Miao L, et al. Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-α. Cell. 2009;137(6):1100-1111.
- Holler N, Zaru R, Micheau O, et al. Fas triggers an alternative, caspase-8–independent cell death pathway using the kinase RIP as effector molecule. Nature immunology. 2000;1(6):489-495.
- Murphy JM, Czabotar PE, Hildebrand JM, et al. The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. J Immunity. 2013;39(3):443-453.
- Sun L, Wang H, Wang Z, et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell. 2012;148(1-2):213-227.
- Zhang D-W, Shao J, Lin J, et al. RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science. 2009;325(5938):332-336.
- Petrie EJ, Hildebrand JM, Murphy JM. Insane in the membrane: a structural perspective of MLKL function in necroptosis. Immunology cell biology. 2017;95(2):152-159.
- Li Y, Wang Z, Jin J, et al. Quercetin pretreatment enhances the radiosensitivity of colon cancer cells by targeting Notch-1 pathway. Biochem Biophys Res Commun. Mar 19 2020;523(4):947-953. doi:10.1016/j.bbrc.2020.01.048
- Kim GT, Lee SH, Kim YM. Quercetin Regulates Sestrin 2-AMPK-mTOR Signaling Pathway and Induces Apoptosis via Increased Intracellular ROS in HCT116 Colon Cancer Cells. J Cancer Prev. Sep 2013;18(3):264-70. doi:10.15430/jcp.2013.18.3.264
- Yüksel TN, Bozgeyik E, Yayla M. The effect of quercetin and quercetin-3-d-xyloside on breast cancer proliferation and migration. Journal of Basic Clinical Health Sciences. 2022;6(2):569-578.
- Bozgeyik E, Bagis H, Bozgeyik I, Kocahan S. The roles of long non-coding RNAs in the necroptotic signaling of colon cancer cells. Molecular biology reports. Apr 25 2023;doi:10.1007/s11033-023-08441-1
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. methods. 2001;25(4):402-408.
- Moriwaki K, Bertin J, Gough P, Orlowski G, Chan FKJCd, disease. Differential roles of RIPK1 and RIPK3 in TNF-induced necroptosis and chemotherapeutic agent-induced cell death. 2015;6(2):e1636-e1636.
- Yan J, Wan P, Choksi S, Liu ZG. Necroptosis and tumor progression. Trends in cancer. Jan 2022;8(1):21-27. doi:10.1016/j.trecan.2021.09.003
- Yang H, Xu S, Tang L, et al. Targeting of non-apoptotic cancer cell death mechanisms by quercetin: Implications in cancer therapy. Frontiers in pharmacology. 2022;13:1043056. doi:10.3389/fphar.2022.1043056
- Estrada-Villaseñor E, Delgado-Cedillo A, Hernández-Pérez A, et al. Ultrastructural changes in giant cell tumor of bone cultured cells exposed to quercetin. Ultrastructural pathology. Nov 2 2021;45(6):335-345. doi:10.1080/01913123.2021.1979704
- Khorsandi L, Orazizadeh M, Niazvand F, Abbaspour MR, Mansouri E, Khodadadi A. Quercetin induces apoptosis and necroptosis in MCF-7 breast cancer cells. Bratislavske lekarske listy. 2017;118(2):123-128. doi:10.4149/bll_2017_025
- Fan H, Tang H-B, Shan L-Q, et al. Quercetin prevents necroptosis of oligodendrocytes by inhibiting macrophages/microglia polarization to M1 phenotype after spinal cord injury in rats. 2019;16(1):1-15.
- Liu L, Liu Y, Cheng X, Qiao X. The Alleviative Effects of Quercetin on Cadmium-Induced Necroptosis via Inhibition ROS/iNOS/NF-κB Pathway in the Chicken Brain. Biol Trace Elem Res. Apr 2021;199(4):1584-1594. doi:10.1007/s12011-020-02563-4
The investigation of the effect of quercetin on HT-29 and HCT-116 colon cancer cell lines through the expression of RIPK1, RIPK3, and MLKL genes
Abstract
Aim: Quercetin is a compound with anti-cancer activity in many types of cancer. However, studies showing the effects of quercetin on the necroptosis pathway are limited, and therefore, the aim of this study was to determine the effect of quercetin on the necroptosis pathway.
Materials and Methods: Colon cancer cells were cultured and effect of different concentrations of quercetin on cell viability was determined by MTT method. Afterwards, the expression level of RIPK1, RIPK3 and MLKL genes were analyzed to determine the effect of quercetin on necroptosis.
Results: The active dose of quercetin was found to be 50 µM in HT-29 cells (p=0.0286), while 100 µM was found in HCT-116 cells (p=0.009). A significant increase in the expression level of necroptosis markers was detected in HT-29 cells treated with 50 and 100 µM quercetin.
Conclusion: The results of this study showed that quercetin may be an active regulator of the necroptosis pathway.
Keywords
Ethical Statement
Bu çalışma için etik kurul onayı etik kurul izni alınmamıştır.
Supporting Institution
Bu çalışma, 2209-A Üniversite Öğrencileri Araştırma Projeleri Destekleme Programı kapsamında Adıyaman Üniversitesi, Sağlık Hizmetleri Meslek Yüksekokulu, Tıbbi Laboratuvar Teknikleri Programı öğrencisi Alp Can TUNCER’in 1919B012202294 numaralı projesi ile Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK) tarafından desteklenmiştir.
Project Number
1919B012202294
References
- Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2021;71(3):209-249.
- Darband SG, Kaviani M, Yousefi B, et al. Quercetin: A functional dietary flavonoid with potential chemo-preventive properties in colorectal cancer. J Cell Physiol. Sep 2018;233(9):6544-6560. doi:10.1002/jcp.26595
- Aoyama N, Kawado M, Yamada H, et al. Low intake of vegetables and fruits and risk of colorectal cancer: the Japan Collaborative Cohort Study. Journal of epidemiology. 2014;24(5):353-360.
- Turati F, Rossi M, Pelucchi C, Levi F, La Vecchia C. Fruit and vegetables and cancer risk: a review of southern European studies. British Journal of Nutrition. 2015;113(S2):S102-S110.
- Sak K. Site-specific anticancer effects of dietary flavonoid quercetin. Nutrition cancer. 2014;66(2):177-193.
- van Erk MJ, Roepman P, van der Lende TR, et al. Integrated assessment by multiple gene expression analysis of quercetin bioactivity on anticancer–related mechanisms in colon cancer cells in vitro. European journal of nutrition. 2005;44(3):143-156.
- Kim H-J, Kim S-K, Kim B-S, et al. Apoptotic effect of quercetin on HT-29 colon cancer cells via the AMPK signaling pathway. Journal of agricultural food chemistry. 2010;58(15):8643-8650.
- Wenzel U, Herzog A, Kuntz S, Daniel H. Protein expression profiling identifies molecular targets of quercetin as a major dietary flavonoid in human colon cancer cells. Proteomics. 2004;4(7):2160-2174.
- Kim H-S, Wannatung T, Lee S, et al. Quercetin enhances hypoxia-mediated apoptosis via direct inhibition of AMPK activity in HCT116 colon cancer. Apoptosis. 2012;17(9):938-949.
- Zhang H, Zhang M, Yu L, Zhao Y, He N, Yang X. Antitumor activities of quercetin and quercetin-5′, 8-disulfonate in human colon and breast cancer cell lines. Food Chemical Toxicology. 2012;50(5):1589-1599.
- He S, Wang L, Miao L, et al. Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-α. Cell. 2009;137(6):1100-1111.
- Holler N, Zaru R, Micheau O, et al. Fas triggers an alternative, caspase-8–independent cell death pathway using the kinase RIP as effector molecule. Nature immunology. 2000;1(6):489-495.
- Murphy JM, Czabotar PE, Hildebrand JM, et al. The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. J Immunity. 2013;39(3):443-453.
- Sun L, Wang H, Wang Z, et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell. 2012;148(1-2):213-227.
- Zhang D-W, Shao J, Lin J, et al. RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science. 2009;325(5938):332-336.
- Petrie EJ, Hildebrand JM, Murphy JM. Insane in the membrane: a structural perspective of MLKL function in necroptosis. Immunology cell biology. 2017;95(2):152-159.
- Li Y, Wang Z, Jin J, et al. Quercetin pretreatment enhances the radiosensitivity of colon cancer cells by targeting Notch-1 pathway. Biochem Biophys Res Commun. Mar 19 2020;523(4):947-953. doi:10.1016/j.bbrc.2020.01.048
- Kim GT, Lee SH, Kim YM. Quercetin Regulates Sestrin 2-AMPK-mTOR Signaling Pathway and Induces Apoptosis via Increased Intracellular ROS in HCT116 Colon Cancer Cells. J Cancer Prev. Sep 2013;18(3):264-70. doi:10.15430/jcp.2013.18.3.264
- Yüksel TN, Bozgeyik E, Yayla M. The effect of quercetin and quercetin-3-d-xyloside on breast cancer proliferation and migration. Journal of Basic Clinical Health Sciences. 2022;6(2):569-578.
- Bozgeyik E, Bagis H, Bozgeyik I, Kocahan S. The roles of long non-coding RNAs in the necroptotic signaling of colon cancer cells. Molecular biology reports. Apr 25 2023;doi:10.1007/s11033-023-08441-1
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. methods. 2001;25(4):402-408.
- Moriwaki K, Bertin J, Gough P, Orlowski G, Chan FKJCd, disease. Differential roles of RIPK1 and RIPK3 in TNF-induced necroptosis and chemotherapeutic agent-induced cell death. 2015;6(2):e1636-e1636.
- Yan J, Wan P, Choksi S, Liu ZG. Necroptosis and tumor progression. Trends in cancer. Jan 2022;8(1):21-27. doi:10.1016/j.trecan.2021.09.003
- Yang H, Xu S, Tang L, et al. Targeting of non-apoptotic cancer cell death mechanisms by quercetin: Implications in cancer therapy. Frontiers in pharmacology. 2022;13:1043056. doi:10.3389/fphar.2022.1043056
- Estrada-Villaseñor E, Delgado-Cedillo A, Hernández-Pérez A, et al. Ultrastructural changes in giant cell tumor of bone cultured cells exposed to quercetin. Ultrastructural pathology. Nov 2 2021;45(6):335-345. doi:10.1080/01913123.2021.1979704
- Khorsandi L, Orazizadeh M, Niazvand F, Abbaspour MR, Mansouri E, Khodadadi A. Quercetin induces apoptosis and necroptosis in MCF-7 breast cancer cells. Bratislavske lekarske listy. 2017;118(2):123-128. doi:10.4149/bll_2017_025
- Fan H, Tang H-B, Shan L-Q, et al. Quercetin prevents necroptosis of oligodendrocytes by inhibiting macrophages/microglia polarization to M1 phenotype after spinal cord injury in rats. 2019;16(1):1-15.
- Liu L, Liu Y, Cheng X, Qiao X. The Alleviative Effects of Quercetin on Cadmium-Induced Necroptosis via Inhibition ROS/iNOS/NF-κB Pathway in the Chicken Brain. Biol Trace Elem Res. Apr 2021;199(4):1584-1594. doi:10.1007/s12011-020-02563-4
There are 28 citations in total.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Cancer Cell Biology, Cancer Therapy (Excl. Chemotherapy and Radiation Therapy) |
| Journal Section | Research Article |
| Authors | |
| Project Number | 1919B012202294 |
| Publication Date | December 31, 2023 |
| Submission Date | May 26, 2023 |
| Acceptance Date | September 14, 2023 |
| Published in Issue | Year 2023 Volume: 9 Issue: 3 |
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