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Papain Induces Apoptosis by Regulating Caspase-3 and Caspase-9 Genes in HepG2 Cells

Yıl 2023, Cilt: 45 Sayı: 1, 94 - 100, 23.01.2023
https://doi.org/10.20515/otd.1181399

Öz

Hepatocellular carcinoma (HCC) ranks second in cancer-related death and its incidence is increasing worldwide. Papain is a proteolytic enzyme and a potential anticancer agent. The aim of the study is to evaluate the cytotoxic effect of papain in human hepatoma HepG2 cell line through apoptosis. The cytotoxicity of papain was determined by the MTT method. Morphological changes of Papain-treated HepG2 cells were evaluated by acridine orange and ethidium bromide (AO/EB) dual staining. Apoptotic activity was determined by the expression of apoptosis regulator caspase-3 and caspase-9 genes by qPCR method. As a result of the administration of 500 µg/ml, 250 µg/ml, 100 µg/ml, 50 µg/ml, 25 µg/ml and 10 µg/ml papain to HepG2 cells for 48 hours, the IC50 value was found to be 53 µg/ml. Caspase 3 and 9 gene expressions, which are apoptotic markers, were significantly increased as a result of papain administration in HepG2 cells. In conclusion, papain may have anticancer effect for HCC treatment by inducing expression of apoptosis regulatory genes.

Kaynakça

  • 1. Ahmed DE, Rashidi FB, Abdelhakim HK, ve ark. An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart. Journal of the Egyptian National Cancer Institute. 2021;33(1):1-15.
  • 2. Al-Fatlawi AA, Al-Fatlawi AA, Irshad M, ve ark. Rice bran phytic acid induced apoptosis through regulation of Bcl-2/Bax and p53 genes in HepG2 human hepatocellular carcinoma cells. Asian Pacific Journal of Cancer Prevention. 2014;15(8):3731-6.
  • 3. Ezhilarasan D. Herbal therapy for cancer. In: Timiri Shanmugam P, ed. Understanding Cancer Therapies. Boca Raton: CRC Press; 2018:129-66.
  • 4. Li Y, Martin RC 2nd. Herbal medicine and hepatocellular carcinoma: applications and challenges. Evid Based Complement Alternat Med. 2011;2011:541209.
  • 5. Thakur RS, Devaraj E. Lagerstroemia speciosa (L.) Pers. triggers oxidative stress mediated apoptosis via intrinsic mitochondrial pathway in HepG2 cells. Environmental Toxicology. 2020;35(11):1225-33.
  • 6. Tsuge HT, Nishimura Y, Tada T, ve ark. Inhibition mechanism of cathepsin L-specific inhibitors based on the crystal structure of papain-CLIK148 complex. Biochem. Biophys. Res. Commun. 1999;266:411-6.
  • 7. Mamboya EAF. Papain, a plant enzyme of biological importance: a review. American Journal of Biochemistry and biotechnology. 2012;8(2):99-104.
  • 8. Flindt ML. Allergy to alpha-amylase and papain. Lancet. 1979;1:1407-8.
  • 9. Li T, Yan G, Bai Y, ve ark. Papain bioinspired gold nanoparticles augmented the anticancer potency of 5-FU against lung cancer. Journal of Experimental Nanoscience. 2020;15(1):109-28.
  • 10. Gebauer F, Micheel B, Stauder G, ve ark. Proteolytic enzymes modulate the adhesion molecule CD44 on malignant cells in vitro. Int J Immunother. 1997;13:111–9.
  • 11. Aslan, A. Ratlarda azoksimetan uygulanarak oluşturulan kolorektal kanserde likopenin siklooksijenaz-2 (cox-2), kaspaz-3, kaspaz-9, bax, bcl-2, p53 proteinlerinin ekspresyonu ve DNA hasarı üzerine etkisi/The effect of lycopene on the cyclooxygenase (cox-2), caspase-3, caspase-9, bax, bcl-2, p53 protein expression and DNA damage in rats with azoxhymethane-induced colorectal cancer. Doktora tezi, 2011.
  • 12. Abedin MJ, Wang D, McDonnell MA, ve ark. Autophagy Delays Apoptotic Death in Breast Cancer Cells Following DNA Damage, Cell Death and Differentiation, 2007;14:500-10.
  • 13. Yerlikaya A, Erin N. Differential sensitivity of breast cancer and melanoma cells to proteasome inhibitor Velcade. International Journal of Molecular Medicine. 2008;22(6):817–23.
  • 14. Famta P, Mishra V, Khatik GL. Formulation and evaluation of 5-fluorouracil and methotrexate gold nanoparticles [doctoral dissertation]. Phagwara (India): Lovely Professional University; 2017.
  • 15. Öner Ç, İsan H, Gülhan Aktaş R, ve ark. The Effect of Vitamin D on Hepatocellülar Carcinoma. Osmangazi Journal of Medicine. 2020;301-10.
  • 16. Kuş G. Hepatosellüler Karsinom Hücrelerinde Karmofurun Sitotoksik Ve Apoptotik Etkileri. Kocatepe Medical Journal. 2017:55-60.
  • 17. Xu S, Shen Y, Xu J, ve ark. Antioxidant and anticancer effects in human hepatocarcinoma (HepG2) cells of papain-hydrolyzed sorghum kafirin hydrolysates. Journal of functional foods. 2019;58:374-82.
  • 18. Akila M, Sushama A, Ramanathan K. Study on in vitro cytotoxicity of papain against liver cancer cell line Hep G2. Cell. 2014;8(7.8): 84-12.
  • 19. Mansour GH, El-Magd MA, Mahfouz DH, ve ark. Bee venom and its active component Melittin synergistically potentiate the anticancer effect of Sorafenib against HepG2 cells. Bioorganic Chemistry. 2021;116:105329.

Papain HepG2 Hücrelerinde Kaspaz-3 ve Kaspaz-9 Genlerini Düzenleyerek Apoptozu İndükler

Yıl 2023, Cilt: 45 Sayı: 1, 94 - 100, 23.01.2023
https://doi.org/10.20515/otd.1181399

Öz

Hepatosellüler karsinom (HCC), kansere bağlı ölüm sıralamasında ikinci sırada yer alır ve dünya genelinde sıklığı artmaktadır. Papain proteolitik bir enzimdir ve potansiyel antikanser ajanıdır. Çalışmanın amacı, papainin insan hepatom HepG2 hücre hattındaki sitotoksik etkisinin apoptoz üzerinden değerlendirilmesidir. Papainin sitotoksisitesi MTT yöntemi ile belirlendi. Papain uygulanan HepG2 hücrelerinin morfolojik değişiklikleri, akridin portakalı ve etidyum bromür (AO/EB) ikili boyaması ile değerlendirildi. Apoptotik aktivite qPCR yöntemi ile apoptoz düzenleyici kaspaz-3 ve kaspaz-9 genlerinin anlatımları ile tespit edildi. HepG2 hücrelerine 48 saat boyunca 500 µg/ml, 250 µg/ml, 100 µg/ml, 50 µg/ml, 25 µg/ml ve 10 µg/ml papain uygulaması sonucunda IC50 değeri 53 µg/ml bulundu. Apoptotik belirteçler olan kaspaz 3 ve 9 gen ifadeleri, HepG2 hücrelerinde papain uygulaması sonucunda önemli ölçüde arttı. Sonuç olarak, papain HCC tedavisi için, apoptoz düzenleyici genlerin anlatımını indükleyerek antikanser etkiye sahip olabilir.

Kaynakça

  • 1. Ahmed DE, Rashidi FB, Abdelhakim HK, ve ark. An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart. Journal of the Egyptian National Cancer Institute. 2021;33(1):1-15.
  • 2. Al-Fatlawi AA, Al-Fatlawi AA, Irshad M, ve ark. Rice bran phytic acid induced apoptosis through regulation of Bcl-2/Bax and p53 genes in HepG2 human hepatocellular carcinoma cells. Asian Pacific Journal of Cancer Prevention. 2014;15(8):3731-6.
  • 3. Ezhilarasan D. Herbal therapy for cancer. In: Timiri Shanmugam P, ed. Understanding Cancer Therapies. Boca Raton: CRC Press; 2018:129-66.
  • 4. Li Y, Martin RC 2nd. Herbal medicine and hepatocellular carcinoma: applications and challenges. Evid Based Complement Alternat Med. 2011;2011:541209.
  • 5. Thakur RS, Devaraj E. Lagerstroemia speciosa (L.) Pers. triggers oxidative stress mediated apoptosis via intrinsic mitochondrial pathway in HepG2 cells. Environmental Toxicology. 2020;35(11):1225-33.
  • 6. Tsuge HT, Nishimura Y, Tada T, ve ark. Inhibition mechanism of cathepsin L-specific inhibitors based on the crystal structure of papain-CLIK148 complex. Biochem. Biophys. Res. Commun. 1999;266:411-6.
  • 7. Mamboya EAF. Papain, a plant enzyme of biological importance: a review. American Journal of Biochemistry and biotechnology. 2012;8(2):99-104.
  • 8. Flindt ML. Allergy to alpha-amylase and papain. Lancet. 1979;1:1407-8.
  • 9. Li T, Yan G, Bai Y, ve ark. Papain bioinspired gold nanoparticles augmented the anticancer potency of 5-FU against lung cancer. Journal of Experimental Nanoscience. 2020;15(1):109-28.
  • 10. Gebauer F, Micheel B, Stauder G, ve ark. Proteolytic enzymes modulate the adhesion molecule CD44 on malignant cells in vitro. Int J Immunother. 1997;13:111–9.
  • 11. Aslan, A. Ratlarda azoksimetan uygulanarak oluşturulan kolorektal kanserde likopenin siklooksijenaz-2 (cox-2), kaspaz-3, kaspaz-9, bax, bcl-2, p53 proteinlerinin ekspresyonu ve DNA hasarı üzerine etkisi/The effect of lycopene on the cyclooxygenase (cox-2), caspase-3, caspase-9, bax, bcl-2, p53 protein expression and DNA damage in rats with azoxhymethane-induced colorectal cancer. Doktora tezi, 2011.
  • 12. Abedin MJ, Wang D, McDonnell MA, ve ark. Autophagy Delays Apoptotic Death in Breast Cancer Cells Following DNA Damage, Cell Death and Differentiation, 2007;14:500-10.
  • 13. Yerlikaya A, Erin N. Differential sensitivity of breast cancer and melanoma cells to proteasome inhibitor Velcade. International Journal of Molecular Medicine. 2008;22(6):817–23.
  • 14. Famta P, Mishra V, Khatik GL. Formulation and evaluation of 5-fluorouracil and methotrexate gold nanoparticles [doctoral dissertation]. Phagwara (India): Lovely Professional University; 2017.
  • 15. Öner Ç, İsan H, Gülhan Aktaş R, ve ark. The Effect of Vitamin D on Hepatocellülar Carcinoma. Osmangazi Journal of Medicine. 2020;301-10.
  • 16. Kuş G. Hepatosellüler Karsinom Hücrelerinde Karmofurun Sitotoksik Ve Apoptotik Etkileri. Kocatepe Medical Journal. 2017:55-60.
  • 17. Xu S, Shen Y, Xu J, ve ark. Antioxidant and anticancer effects in human hepatocarcinoma (HepG2) cells of papain-hydrolyzed sorghum kafirin hydrolysates. Journal of functional foods. 2019;58:374-82.
  • 18. Akila M, Sushama A, Ramanathan K. Study on in vitro cytotoxicity of papain against liver cancer cell line Hep G2. Cell. 2014;8(7.8): 84-12.
  • 19. Mansour GH, El-Magd MA, Mahfouz DH, ve ark. Bee venom and its active component Melittin synergistically potentiate the anticancer effect of Sorafenib against HepG2 cells. Bioorganic Chemistry. 2021;116:105329.
Toplam 19 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Sağlık Kurumları Yönetimi
Bölüm ORİJİNAL MAKALELER / ORIGINAL ARTICLES
Yazarlar

Meliha Koldemir Gündüz 0000-0003-0969-6377

Fatih Kar 0000-0001-8356-9806

Güllü Kaymak 0000-0001-6309-0208

Yayımlanma Tarihi 23 Ocak 2023
Yayımlandığı Sayı Yıl 2023 Cilt: 45 Sayı: 1

Kaynak Göster

Vancouver Koldemir Gündüz M, Kar F, Kaymak G. Papain HepG2 Hücrelerinde Kaspaz-3 ve Kaspaz-9 Genlerini Düzenleyerek Apoptozu İndükler. Osmangazi Tıp Dergisi. 2023;45(1):94-100.


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