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Keçiboynuzu ekstrelerinin insan servikal kanser hücreleri üzerindeki antiproliferatif ve sitotoksik etkileri

Year 2018, Volume: 15 Issue: 3, 199 - 206, 12.12.2018

Abstract

Amaç: Leguminosae familyasının bir üyesi olan Ceratonia siliqua L., (keçiboynuzu, harnup)’nın antimikrobiyal, antifungal ve sitotoksik aktiviteye sahip olduğu bilinmektedir. Bu çalışmada, keçiboynuzunun yaprak ve meyvelerinin etli kısımlarından metanol ve etanol ile hazırlanan ekstrelerin, HeLa hücreleri üzerindeki antiproliferatif ve sitotoksik etkileri araştırılmıştır. Ayrıca embriyonik fare fibroblast hücre soyu olan 3T3 hücreleri kontrol olarak kullanılmıştır.

Materyal ve Metod: Bu amaçlarla 48 saat süreyle ve farklı konsantrasyonlarda (200-800μg/mL) ekstre uygulamasının ardından canlı hücre sayıları (proliferasyon, MTT yöntemi), mitotik indeks (mitotik aktivite) ve işaretlenme indeksleri (DNA sentezi) tespit edilmiştir. Ayrıca ekstrelerde bulunan fenolik bileşen kompozisyonları da LC-MS/MS ile belirlenmiştir.

Bulgular: Elde edilen bulgulara göre keçiboynuzunun metanolik ve etanolik yaprak ekstreleri insan servikal kanser HeLa hücrelerinin DNA sentezini inhibe ederek mitotik aktivitesini ve proliferasyonunu baskılamıştır. Ancak bu ekstreler aynı zamanda normal fibroblast 3T3 hücrelerine karşı da sitotoksik etki göstermişlerdir. Etanolik meyve ekstresi HeLa hücrelerinin DNA sentezini azaltmıştır.

Sonuç: C. siliqua bitkisinden elde edilecek etken maddeler veya hazırlanacak preparasyonlar antikanser ajan olabilme potansiyeli taşıyabilir. Ancak normal hücreler üzerindeki sitotoksik etkiler de göz ardı edilmemelidir.

References

  • 1. Siegel RL, Miller KB, Jemal A. Cancer Statistic. CA Cancer J Clin 2017;67:7-30.
  • 2. Kayaalp SO. Tıbbi Farmakoloji. Ankara: Hacettepe-Taş Kitapçılık. 1996.
  • 3. Weaver BA. How Taxol/paclitaxel kills cancer cells. Mol Biol Cell 2014;25:2677-2681.
  • 4. Thirumaran R, Prendergast GC., Gilman PB. Cytotoxic Chemotherapy in Clinical Treatment of Cancer. In: Prendergast GC., Jaffee EM. eds. Cancer Immunotherapy. Academic Press. 2007:101-116.
  • 5. Seçmen Ö, Gemici Y, Görk G, Bekat L, Leblebici E. Tohumlu Bitkiler Sistematiği, 4. Baskı, İzmir:Ege Üniversitesi Basımevi 1995.
  • 6. Baytop, T. Türkiye’de Bitkilerle Tedavi Geçmişte ve Bugün. 2. Baskı. İstanbul: Nobel Tıp Kitabevleri. 1999.
  • 7. Kıvçak B, Mert T, Öztürk T. Antimicrobial and cytotoxic activities of Ceratonia siliqua L. extracts. Turk J Biol 2002;26:197-200.
  • 8. Corsi L, Avallone R, Cosenza F, Farina F, Baraldi C, Baraldi M. Antiproliferative effects of Ceratonia siliqua L. on mouse hepatocellular carcinoma cell line. Fitoterapia 2002;73:674-684.
  • 9. Mosmann T. Rapid Colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
  • 10. Bancroft JD, Gamble M. Theory and Practice of Histological Techniques. 3rd Edn. London: Elsevier, 1990.
  • 11. Özalpan A. Temel Radyobiyoloji, 1. Baskı, İstanbul: Haliç Üniversitesi Yayınları. 2001.
  • 12. Avallone R, Cosenza F, Farina F, Baraldi C, Baraldi M. Extraction and purification from Ceratonia siliqua of compounds acting on central and peripheral benzodiazepine receptors, Fitoterapia 2002;73:390-396.
  • 13. Klenow S, Glei M, Haber B, Owen R, Pool-Zobel BL. Carob fibre compounds modulate parameters of cell growth differently in human HT29 colon adenocarcinoma cells than in LT97 colon adenoma cells, Food Chem Toxicol 2008;46:1389-1397.
  • 14. Owen RW, Haubner R, Hull WE, ve ark. Isolation and structure elucidation of major individual polyphenols in carob fibre. Food Chem Toxicol 2003;41:1727-1738.
  • 15. De Ancos B, Gonzalez EM, Cano MP. Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit, J Agric Food Chem 2000;48:4565-4570.
  • 16. Patel AV, Rojas-Vera J, Dacke CG. Therapeutic constituents and actions of Rubus species, Curr Med Chem 2004;11:1501-1512.
  • 17. Custódio L, Escapa AL, Fernandes E, Fajardo A, Aligué R, Albericio F, Neng N, Nogueira JMF, Romano A. Phytochemical profile, antioxidant and cytotoxic activities of the carob tree (Ceratonia siliqua L.) germ flour extracts. Plant Foods Hum Nutr 2011;66:78–84.
  • 18. Custódio L, Fernandes E, Escapa AL, Fajardo A, Aligué R, Albericio F, Neng N, Nogueira JMF, Romano A. Antioxidant and cytotoxic activities of carob tree fruit pulps are strongly ınfluenced by gender and cultivar. J Agric Food Chem 2011;59:7005–7012.
  • 19. Paterson JR, Lawrence JR. Salicylic acid: a link between aspirin, diet and the prevention of colorectal cancer. QJ Med 2001;94:445-448.
  • 20. Yang J-H, Hsia T-C, Kuo H-M, Lee Chao P-D, Chou C-C, Wei Y-H, Chung J-G. Inhibition of lung cancer cell growth by quercetin glucuronides via G2/M arrest and induction of apoptosis, Drug Metab Dispos 2006;34(2):296-304.
  • 21. Spitz GA, Furtado CM, Sola-Penna M, Zancan P. Acetylsalicylic acid and salicylic acid decrease tumor cell viability and glucose metabolism modulating 6-phosphofructo-1-kinase structure and activity. Biochem Pharmacol 2009;77:46–53.
  • 22. Watson RR, Preedy VR, Zibadi S. eds. Polyphenols in Human Health and Disease, USA:Elsevier, 2014.
  • 23. Cook MT. Mechanism of metastasis suppression by luteolin in breast cancer. Breast Cancer 2018;10:89–100.
  • 24. Han K, Lang T, Zhang Z, Zhang Y, Sun Y, Shen Z, Beuerman RW, Zhou L, Min D. Luteolin attenuates Wnt signaling via upregulation of FZD6 to suppress prostate cancer stemness revealed by comparative proteomics. Sci Rep 2018;8:8537.
  • 25. Chen P-Y, Tien H-J, Chen S-F, Horng C-T, Tang H-L, Jung H-L, Wu M-J, Yen J-H. Response of myeloid leukemia cells to luteolin is modulated by differentially expressed Pituitary Tumor-Transforming Gene 1 (PTTG1) oncoprotein. Int J Mol Sci 2018;19:1173.
  • 26. Custódio L, Fernandes E, Escapa AL, ve ark. Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua), Pharm Biol 2009;47(8):721-728.

Antiproliferative and cytotoxic effects of carob extracts on human cervical cancer cells

Year 2018, Volume: 15 Issue: 3, 199 - 206, 12.12.2018

Abstract

Background: Ceratonia siliqua L., a member of the family Leguminosae, has been reported to have antimicrobial, antifungal and cytotoxic activity (carob, kharnub). In this study, the antiproliferative and cytotoxic effects of extracts prepared with methanol and ethanol from the carob leaves and fruits on HeLa cells were investigated. Additionally, 3T3, embryonic mouse fibroblast cell line, was used as a control.

Material and Methods: For these purposes, after treatment of the extracts at different concentrations (200-800μg/mL) for 48 hours, viable cell counts (proliferation, MTT method), mitotic index (mitotic activity) and labeling index (DNA synthesis) were determined. In addition, the phenolic component compositions in the extracts were also determined by LC-MS / MS.

Results: According to the findings, the methanolic and ethanolic leaf extracts of carob bean suppressed mitotic activity and proliferation by inhibiting the DNA synthesis of human cervical cancer HeLa cells. However, these extracts also exhibited a cytotoxic effect against normal fibroblast 3T3 cells. Ethanolic fruit extract reduced the DNA synthesis of HeLa cells.

Conclusions: The active substances or preparations obtaining from the C. siliqua plant have the possibility of being an anticancer agent, whereas their cytotoxic effects on the normal cells should not be overlooked.

References

  • 1. Siegel RL, Miller KB, Jemal A. Cancer Statistic. CA Cancer J Clin 2017;67:7-30.
  • 2. Kayaalp SO. Tıbbi Farmakoloji. Ankara: Hacettepe-Taş Kitapçılık. 1996.
  • 3. Weaver BA. How Taxol/paclitaxel kills cancer cells. Mol Biol Cell 2014;25:2677-2681.
  • 4. Thirumaran R, Prendergast GC., Gilman PB. Cytotoxic Chemotherapy in Clinical Treatment of Cancer. In: Prendergast GC., Jaffee EM. eds. Cancer Immunotherapy. Academic Press. 2007:101-116.
  • 5. Seçmen Ö, Gemici Y, Görk G, Bekat L, Leblebici E. Tohumlu Bitkiler Sistematiği, 4. Baskı, İzmir:Ege Üniversitesi Basımevi 1995.
  • 6. Baytop, T. Türkiye’de Bitkilerle Tedavi Geçmişte ve Bugün. 2. Baskı. İstanbul: Nobel Tıp Kitabevleri. 1999.
  • 7. Kıvçak B, Mert T, Öztürk T. Antimicrobial and cytotoxic activities of Ceratonia siliqua L. extracts. Turk J Biol 2002;26:197-200.
  • 8. Corsi L, Avallone R, Cosenza F, Farina F, Baraldi C, Baraldi M. Antiproliferative effects of Ceratonia siliqua L. on mouse hepatocellular carcinoma cell line. Fitoterapia 2002;73:674-684.
  • 9. Mosmann T. Rapid Colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
  • 10. Bancroft JD, Gamble M. Theory and Practice of Histological Techniques. 3rd Edn. London: Elsevier, 1990.
  • 11. Özalpan A. Temel Radyobiyoloji, 1. Baskı, İstanbul: Haliç Üniversitesi Yayınları. 2001.
  • 12. Avallone R, Cosenza F, Farina F, Baraldi C, Baraldi M. Extraction and purification from Ceratonia siliqua of compounds acting on central and peripheral benzodiazepine receptors, Fitoterapia 2002;73:390-396.
  • 13. Klenow S, Glei M, Haber B, Owen R, Pool-Zobel BL. Carob fibre compounds modulate parameters of cell growth differently in human HT29 colon adenocarcinoma cells than in LT97 colon adenoma cells, Food Chem Toxicol 2008;46:1389-1397.
  • 14. Owen RW, Haubner R, Hull WE, ve ark. Isolation and structure elucidation of major individual polyphenols in carob fibre. Food Chem Toxicol 2003;41:1727-1738.
  • 15. De Ancos B, Gonzalez EM, Cano MP. Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit, J Agric Food Chem 2000;48:4565-4570.
  • 16. Patel AV, Rojas-Vera J, Dacke CG. Therapeutic constituents and actions of Rubus species, Curr Med Chem 2004;11:1501-1512.
  • 17. Custódio L, Escapa AL, Fernandes E, Fajardo A, Aligué R, Albericio F, Neng N, Nogueira JMF, Romano A. Phytochemical profile, antioxidant and cytotoxic activities of the carob tree (Ceratonia siliqua L.) germ flour extracts. Plant Foods Hum Nutr 2011;66:78–84.
  • 18. Custódio L, Fernandes E, Escapa AL, Fajardo A, Aligué R, Albericio F, Neng N, Nogueira JMF, Romano A. Antioxidant and cytotoxic activities of carob tree fruit pulps are strongly ınfluenced by gender and cultivar. J Agric Food Chem 2011;59:7005–7012.
  • 19. Paterson JR, Lawrence JR. Salicylic acid: a link between aspirin, diet and the prevention of colorectal cancer. QJ Med 2001;94:445-448.
  • 20. Yang J-H, Hsia T-C, Kuo H-M, Lee Chao P-D, Chou C-C, Wei Y-H, Chung J-G. Inhibition of lung cancer cell growth by quercetin glucuronides via G2/M arrest and induction of apoptosis, Drug Metab Dispos 2006;34(2):296-304.
  • 21. Spitz GA, Furtado CM, Sola-Penna M, Zancan P. Acetylsalicylic acid and salicylic acid decrease tumor cell viability and glucose metabolism modulating 6-phosphofructo-1-kinase structure and activity. Biochem Pharmacol 2009;77:46–53.
  • 22. Watson RR, Preedy VR, Zibadi S. eds. Polyphenols in Human Health and Disease, USA:Elsevier, 2014.
  • 23. Cook MT. Mechanism of metastasis suppression by luteolin in breast cancer. Breast Cancer 2018;10:89–100.
  • 24. Han K, Lang T, Zhang Z, Zhang Y, Sun Y, Shen Z, Beuerman RW, Zhou L, Min D. Luteolin attenuates Wnt signaling via upregulation of FZD6 to suppress prostate cancer stemness revealed by comparative proteomics. Sci Rep 2018;8:8537.
  • 25. Chen P-Y, Tien H-J, Chen S-F, Horng C-T, Tang H-L, Jung H-L, Wu M-J, Yen J-H. Response of myeloid leukemia cells to luteolin is modulated by differentially expressed Pituitary Tumor-Transforming Gene 1 (PTTG1) oncoprotein. Int J Mol Sci 2018;19:1173.
  • 26. Custódio L, Fernandes E, Escapa AL, ve ark. Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua), Pharm Biol 2009;47(8):721-728.
There are 26 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

Hatice Gümüşhan Aktaş 0000-0002-6650-184X

Publication Date December 12, 2018
Submission Date July 22, 2018
Acceptance Date November 6, 2018
Published in Issue Year 2018 Volume: 15 Issue: 3

Cite

Vancouver Gümüşhan Aktaş H. Keçiboynuzu ekstrelerinin insan servikal kanser hücreleri üzerindeki antiproliferatif ve sitotoksik etkileri. Harran Üniversitesi Tıp Fakültesi Dergisi. 2018;15(3):199-206.

Harran Üniversitesi Tıp Fakültesi Dergisi  / Journal of Harran University Medical Faculty