HCT-116 Kolon Kanseri Hücreleri Üzerinde Kannabinolün Sitotoksik ve Metastatik Etkilerinin Değerlendirilmesi
Yıl 2025,
Cilt: 47 Sayı: 4, 593 - 599, 18.06.2025
Gamze Turna Saltoğlu
,
Serap Yalcin
,
Bayram Furkan Coşkun
,
Habibe Sema Arslan
Öz
Kannabinolün (CBN) HCT-116 kolon kanseri hücreleri üzerindeki sitotoksik ve metastatik etkilerini değerlendirmeyi amaçlamaktadır. Kolorektal kanser dünya genelinde kanserle ilişkili ölümlerin en yaygın nedenlerinden biridir ve mevcut tedavi yaklaşımlarına rağmen yüksek nüks ve metastaz oranları görülmektedir. Son yıllarda, kannabinoidlerin antitümöral potansiyeli araştırılmakta olup, CBN'nin bu konudaki etkileri henüz tam olarak aydınlatılmamıştır. HCT-116 kolon kanseri hücreleri laboratuvar koşullarında kültüre edilerek CBN ile muamele edildi. Hücre canlılığı XTT sitotoksisite testi ile,hücre göçü ve invazyon yeteneği sırasıyla yara iyileşme testi (wound healing assay) ve invazyon analizi ile değerlendirildi. Elde edilen veriler, ImageJ yazılımı ile sayısallaştırıldı ve GraphPad Prism 8 programı kullanılarak istatistiksel olarak değerlendirildi. CBN’nin IC₅₀ değeri 69.14 μM olarak belirlendi ve CBN uygulamasının hücre canlılığında anlamlı bir azalmaya neden olduğu belirlendi. (p<0.05). Yara iyileşme testi sonuçları, CBN uygulamasının hücre migrasyonunu belirgin şekilde inhibe ettiğini ortaya koymuştur (p<0.05). İnvazyon analizleri, CBN’nin invaziviteyi belirgin derecede azalttığını ve kontrol grubuna kıyasla anlamlı bir inhibisyon sağladığını göstermektedir (p<0.05). Bu bulgular, CBN’nin HCT-116 hücrelerinde metastatik süreçleri baskıladığına dair önemli kanıtlar sunmaktadır. Bu çalışma CBN’nin kolon kanseri hücreleri üzerinde sitotoksik ve metastatik aktiviteyi inhibe edici potansiyelini desteklemektedir. Bulgular, CBN’nin hücre proliferasyonunu, migrasyonunu ve invazyonunu azalttığını göstermekte olup, kolorektal kanser tedavisinde CBN’nin olası terapötik bir ajan olarak değerlendirilebileceğini düşündürmektedir. Ancak, bu etkinin altta yatan moleküler mekanizmalarının daha ayrıntılı olarak araştırılması ve in vivo modellerde test edilmesi gerekmektedir.
Kaynakça
- 1. Siegel RL, Miller KD, Goding Sauer A, Fedewa SA, Butterly LF, Anderson JC, et al. Colorectal cancer statistics, 2020. CA: a cancer journal for clinicians. 2020;70(3):145-64.
- 2. Ahmed M. Colon cancer: a clinician’s perspective in 2019. Gastroenterology research. 2020;13(1):1.
- 3. Islam MR, Akash S, Rahman MM, Nowrin FT, Akter T, Shohag S, et al. Colon cancer and colorectal cancer: Prevention and treatment by potential natural products. Chemico-biological interactions. 2022;368:110170.
- 4. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, 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):209-49.
- 5. Kuppusamy P, Yusoff MM, Maniam GP, Ichwan SJA, Soundharrajan I, Govindan N. Nutraceuticals as potential therapeutic agents for colon cancer: a review. Acta Pharmaceutica Sinica B. 2014;4(3):173-81.
- 6. Haggar FA, Boushey RP. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clinics in colon and rectal surgery. 2009;22(04):191-7.
- 7. Gonçalves ECD, Baldasso GM, Bicca MA, Paes RS, Capasso R, Dutra RC. Terpenoids, Cannabimimetic Ligands, beyond the Cannabis Plant. Molecules. 2020;25(7):1567.
- 8. Kalant H. Medicinal use of cannabis: history and current status. Pain Research and Management. 2001;6(2):80-91.
- 9. O'Shaughnessy WB. On the preparations of the Indian hemp, or Gunjah: Cannabis indica their effects on the animal system in health, and their utility in the treatment of tetanus and other convulsive diseases. Provincial Medical Journal and Retrospect of the Medical Sciences. 1843;5(123):363.
- 10. Zuardi AW. History of cannabis as a medicine: a review. Brazilian Journal of Psychiatry. 2006;28:153-7.
- 11. Kleckner AS, Kleckner IR, Kamen CS, Tejani MA, Janelsins MC, Morrow GR, et al. Opportunities for cannabis in supportive care in cancer. Therapeutic advances in medical oncology. 2019;11:1758835919866362.
- 12. Parker LA, Rock EM, Limebeer CL. Regulation of nausea and vomiting by cannabinoids. British journal of pharmacology. 2011;163(7):1411-22.
- 13. Pertwee RG. Cannabinoid pharmacology: the first 66 years. British journal of pharmacology. 2006;147(S1):S163-S71.
- 14. Kovalchuk O, Kovalchuk I. Cannabinoids as anticancer therapeutic agents. Cell Cycle. 2020;19(9):961-89.
- 15. Guindon J, Hohmann AG. The endocannabinoid system and pain. CNS & Neurological Disorders-Drug Targets (Formerly Current Drug Targets-CNS & Neurological Disorders). 2009;8(6):403-21.
- 16. Andre CM, Hausman J-F, Guerriero G. Cannabis sativa: the plant of the thousand and one molecules. Frontiers in plant science. 2016;7:19.
- 17. Morales P, Hurst DP, Reggio PH. Molecular Targets of the Phytocannabinoids: A Complex Picture. Prog Chem Org Nat Prod. 2017;103:103-31.
- 18. Velasco G, Sánchez C, Guzmán M. Anticancer Mechanisms of Cannabinoids. Current Oncology. 2016;23(11):23-32.
- 19. Pertwee R. The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: Δ9‐tetrahydrocannabinol, cannabidiol and Δ9‐tetrahydrocannabivarin. British journal of pharmacology. 2008;153(2):199-215.
- 20. Velasco G, Sánchez C, Guzmán M. Towards the use of cannabinoids as antitumour agents. Nature Reviews Cancer. 2012;12(6):436-44.
- 21. Wang B, Li D, Cherkasova V, Gerasymchuk M, Narendran A, Kovalchuk I, et al. Cannabinol Inhibits Cellular Proliferation, Invasion, and Angiogenesis of Neuroblastoma via Novel miR-34a/tRiMetF31/PFKFB3 Axis. Cancers. 2022;14(8):1908.
- 22. Nallathambi R, Mazuz M, Namdar D, Shik M, Namintzer D, Vinayaka AC, et al. Identification of Synergistic Interaction Between Cannabis-Derived Compounds for Cytotoxic Activity in Colorectal Cancer Cell Lines and Colon Polyps That Induces Apoptosis-Related Cell Death and Distinct Gene Expression. Cannabis and Cannabinoid Research. 2018;3(1):120-35.
- 23. Shaw LM. Tumor cell invasion assays. Cell migration: developmental methods and protocols. 2005:97-105.
- 24. Rodriguez LG, Wu X, Guan J-L. Wound-healing assay. Cell Migration: Developmental Methods and Protocols. 2005:23-9.
- 25. Eng C, Yoshino T, Ruíz-García E, Mostafa N, Cann CG, O'Brian B, et al. Colorectal cancer. The Lancet. 2024;404(10449):294-310.
- 26. Wang X, Zhang L, Li H, Sun W, Zhang H, Lai M. THBS2 is a potential prognostic biomarker in colorectal cancer. Scientific reports. 2016;6(1):33366.
- 27. Mahadevan A, Siegel C, Martin BR, Abood ME, Beletskaya I, Razdan RK. Novel cannabinol probes for CB1 and CB2 cannabinoid receptors. Journal of medicinal chemistry. 2000;43(20):3778-85.
- 28. Hwang Y-N, Kwon I-S, Park J-H, Na H-H, Kwon T-H, Park J-S, et al. Cell death induction and intracellular vesicle formation in human colorectal cancer cells treated with Δ9-Tetrahydrocannabinol. Genes & Genomics. 2023;45(12):1463-74.
- 29. Salazar M, Carracedo A, Salanueva ÍJ, Hernández-Tiedra S, Lorente M, Egia A, et al. Cannabinoid action induces autophagy-mediated cell death through stimulation of ER stress in human glioma cells. The Journal of clinical investigation. 2009;119(5):1359-72.
- 30. Vara D, Salazar M, Olea-Herrero N, Guzmán M, Velasco G, Díaz-Laviada I. Anti-tumoral action of cannabinoids on hepatocellular carcinoma: role of AMPK-dependent activation of autophagy. Cell Death & Differentiation. 2011;18(7):1099-111.
- 31. Zhong N, Li D, Wang B, Kovalchuk O, Kovalchuk I. Cannabinol inhibits cell growth and triggers cell cycle arrest and apoptosis in cancer cells. Biocatalysis and Agricultural Biotechnology. 2023;48:102627.
- 32. Yousef B, Hassan H, Guerram M, Hamdi A, Wang B, Zhang L-Y, et al. Pristimerin inhibits proliferation, migration and invasion, and induces apoptosis in HCT-116 colorectal cancer cells. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie. 2016;79:112-9.
Evaluating the Cytotoxic and Metastatic Actions of Cannabinol on HCT-116 Colon Cancer Cells
Yıl 2025,
Cilt: 47 Sayı: 4, 593 - 599, 18.06.2025
Gamze Turna Saltoğlu
,
Serap Yalcin
,
Bayram Furkan Coşkun
,
Habibe Sema Arslan
Öz
It aims to evaluate the cytotoxic and metastatic effects of cannabinol (CBN) on HCT-116 colon cancer cells. Colorectal cancer is one of the leading causes of cancer-related mortality worldwide, with high recurrence and metastasis rates despite current treatment approaches. In recent years, the antitumor potential of cannabinoids has gained attention; however, the specific effects of CBN remain largely unexplored. HCT-116 colon cancer cells were cultured under laboratory conditions and treated with CBN. Cell viability was assessed using the XTT cytotoxicity assay, while cell migration and invasion capabilities were analyzed through the wound healing assay and invasion assay, respectively. The acquired data were quantified using ImageJ software and statistically evaluated with GraphPad Prism 8. The IC₅₀ value of CBN was determined to be 69.14 μM, and CBN treatment was found to significantly reduce cell viability (p<0.05). Wound healing assay results demonstrated that CBN treatment markedly inhibited cell migration (p<0.05). Invasion analyses revealed that CBN significantly reduced invasiveness and provided a notable inhibition compared to the control group (p<0.05). This provides significant evidence that CBN suppresses metastatic processes in HCT-116 cells. This study supports the potential of CBN as an inhibitor of cytotoxic and metastatic activity in colon cancer cells. The findings indicate that CBN reduces cell proliferation, migration, and invasion, suggesting its potential as a therapeutic agent in colorectal cancer treatment. However, further research is needed to elucidate the underlying molecular mechanisms and validate these effects in in vivo models.
Kaynakça
- 1. Siegel RL, Miller KD, Goding Sauer A, Fedewa SA, Butterly LF, Anderson JC, et al. Colorectal cancer statistics, 2020. CA: a cancer journal for clinicians. 2020;70(3):145-64.
- 2. Ahmed M. Colon cancer: a clinician’s perspective in 2019. Gastroenterology research. 2020;13(1):1.
- 3. Islam MR, Akash S, Rahman MM, Nowrin FT, Akter T, Shohag S, et al. Colon cancer and colorectal cancer: Prevention and treatment by potential natural products. Chemico-biological interactions. 2022;368:110170.
- 4. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, 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):209-49.
- 5. Kuppusamy P, Yusoff MM, Maniam GP, Ichwan SJA, Soundharrajan I, Govindan N. Nutraceuticals as potential therapeutic agents for colon cancer: a review. Acta Pharmaceutica Sinica B. 2014;4(3):173-81.
- 6. Haggar FA, Boushey RP. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clinics in colon and rectal surgery. 2009;22(04):191-7.
- 7. Gonçalves ECD, Baldasso GM, Bicca MA, Paes RS, Capasso R, Dutra RC. Terpenoids, Cannabimimetic Ligands, beyond the Cannabis Plant. Molecules. 2020;25(7):1567.
- 8. Kalant H. Medicinal use of cannabis: history and current status. Pain Research and Management. 2001;6(2):80-91.
- 9. O'Shaughnessy WB. On the preparations of the Indian hemp, or Gunjah: Cannabis indica their effects on the animal system in health, and their utility in the treatment of tetanus and other convulsive diseases. Provincial Medical Journal and Retrospect of the Medical Sciences. 1843;5(123):363.
- 10. Zuardi AW. History of cannabis as a medicine: a review. Brazilian Journal of Psychiatry. 2006;28:153-7.
- 11. Kleckner AS, Kleckner IR, Kamen CS, Tejani MA, Janelsins MC, Morrow GR, et al. Opportunities for cannabis in supportive care in cancer. Therapeutic advances in medical oncology. 2019;11:1758835919866362.
- 12. Parker LA, Rock EM, Limebeer CL. Regulation of nausea and vomiting by cannabinoids. British journal of pharmacology. 2011;163(7):1411-22.
- 13. Pertwee RG. Cannabinoid pharmacology: the first 66 years. British journal of pharmacology. 2006;147(S1):S163-S71.
- 14. Kovalchuk O, Kovalchuk I. Cannabinoids as anticancer therapeutic agents. Cell Cycle. 2020;19(9):961-89.
- 15. Guindon J, Hohmann AG. The endocannabinoid system and pain. CNS & Neurological Disorders-Drug Targets (Formerly Current Drug Targets-CNS & Neurological Disorders). 2009;8(6):403-21.
- 16. Andre CM, Hausman J-F, Guerriero G. Cannabis sativa: the plant of the thousand and one molecules. Frontiers in plant science. 2016;7:19.
- 17. Morales P, Hurst DP, Reggio PH. Molecular Targets of the Phytocannabinoids: A Complex Picture. Prog Chem Org Nat Prod. 2017;103:103-31.
- 18. Velasco G, Sánchez C, Guzmán M. Anticancer Mechanisms of Cannabinoids. Current Oncology. 2016;23(11):23-32.
- 19. Pertwee R. The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: Δ9‐tetrahydrocannabinol, cannabidiol and Δ9‐tetrahydrocannabivarin. British journal of pharmacology. 2008;153(2):199-215.
- 20. Velasco G, Sánchez C, Guzmán M. Towards the use of cannabinoids as antitumour agents. Nature Reviews Cancer. 2012;12(6):436-44.
- 21. Wang B, Li D, Cherkasova V, Gerasymchuk M, Narendran A, Kovalchuk I, et al. Cannabinol Inhibits Cellular Proliferation, Invasion, and Angiogenesis of Neuroblastoma via Novel miR-34a/tRiMetF31/PFKFB3 Axis. Cancers. 2022;14(8):1908.
- 22. Nallathambi R, Mazuz M, Namdar D, Shik M, Namintzer D, Vinayaka AC, et al. Identification of Synergistic Interaction Between Cannabis-Derived Compounds for Cytotoxic Activity in Colorectal Cancer Cell Lines and Colon Polyps That Induces Apoptosis-Related Cell Death and Distinct Gene Expression. Cannabis and Cannabinoid Research. 2018;3(1):120-35.
- 23. Shaw LM. Tumor cell invasion assays. Cell migration: developmental methods and protocols. 2005:97-105.
- 24. Rodriguez LG, Wu X, Guan J-L. Wound-healing assay. Cell Migration: Developmental Methods and Protocols. 2005:23-9.
- 25. Eng C, Yoshino T, Ruíz-García E, Mostafa N, Cann CG, O'Brian B, et al. Colorectal cancer. The Lancet. 2024;404(10449):294-310.
- 26. Wang X, Zhang L, Li H, Sun W, Zhang H, Lai M. THBS2 is a potential prognostic biomarker in colorectal cancer. Scientific reports. 2016;6(1):33366.
- 27. Mahadevan A, Siegel C, Martin BR, Abood ME, Beletskaya I, Razdan RK. Novel cannabinol probes for CB1 and CB2 cannabinoid receptors. Journal of medicinal chemistry. 2000;43(20):3778-85.
- 28. Hwang Y-N, Kwon I-S, Park J-H, Na H-H, Kwon T-H, Park J-S, et al. Cell death induction and intracellular vesicle formation in human colorectal cancer cells treated with Δ9-Tetrahydrocannabinol. Genes & Genomics. 2023;45(12):1463-74.
- 29. Salazar M, Carracedo A, Salanueva ÍJ, Hernández-Tiedra S, Lorente M, Egia A, et al. Cannabinoid action induces autophagy-mediated cell death through stimulation of ER stress in human glioma cells. The Journal of clinical investigation. 2009;119(5):1359-72.
- 30. Vara D, Salazar M, Olea-Herrero N, Guzmán M, Velasco G, Díaz-Laviada I. Anti-tumoral action of cannabinoids on hepatocellular carcinoma: role of AMPK-dependent activation of autophagy. Cell Death & Differentiation. 2011;18(7):1099-111.
- 31. Zhong N, Li D, Wang B, Kovalchuk O, Kovalchuk I. Cannabinol inhibits cell growth and triggers cell cycle arrest and apoptosis in cancer cells. Biocatalysis and Agricultural Biotechnology. 2023;48:102627.
- 32. Yousef B, Hassan H, Guerram M, Hamdi A, Wang B, Zhang L-Y, et al. Pristimerin inhibits proliferation, migration and invasion, and induces apoptosis in HCT-116 colorectal cancer cells. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie. 2016;79:112-9.