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İsatinin HepG2 ve AML12 hücre hatları üzerindeki moleküler etkilerinin araştırılması

Year 2024, Volume: 14 Issue: 2, 394 - 401, 15.06.2024
https://doi.org/10.17714/gumusfenbil.1389743

Abstract

İsatin, indol türevi olan organik bir bileşiktir. Couroupita guianensis bitkisinin doğal bir bileşenidir. İnsan vücudu hormonu olan adrenalinin de metabolik türevidir. Yapılan çalışmalarda isatin türevlerinin anti-tümöral etkileri gösterilmiştir. Bu çalışmada isatinin hepatoselüler kanser hücre hattı olan HepG2 üzerindeki sitotoksik etkileri ve sağlıklı karaciğer hücre hattı olan AML12 üzerindeki sitotoksik, koruyucu-proliferatif etkileri MTT, floresan boyama, yara iyileşmesi ve gerçek zamanlı polimeraz zincir reaksiyonu analizleri ile araştırıldı. 48 saatlik isatin uygulaması için IC50 değerleri HepG2 için 186.23 µM, AML12 için 7.05 mM olarak hesaplandı. Her iki hücre hattı için HepG2 uygulamasının IC50 dozuna bağlı olarak, yara iyileşmesi ve floresan boyama analizleri, HepG2 hücrelerinde proliferasyonun baskılandığını ve apoptozun tetiklendiğini, ancak AML12 hücrelerinde proliferasyonun teşvik edildiğini göstermiştir. Ek olarak, oksidatif stres genleri CuZn/Mn-SOD ve mitokondriyal apoptotik yol genleri Bax, kesilmiş-Cas3, APAF1 ve p53'ün ekspresyon seviyeleri HepG2'de artmış ancak AML12 hücre hatlarında azalmıştır. Bu çalışmalar sonucunda isatinin doğal formunun karaciğer kanseri hücre dizileri için iyi bir anti-kanser ajanı ve sağlıklı karaciğer hücreleri için koruyucu bir takviye olabileceği sonucuna varılmıştır.

References

  • Abdoul-Fadl, T., & Bin-Jubair, F.A.S. (2010). Anti-tubercular activity of isatin and derivatives. International Journal of Research in Pharmaceutical Sciences, 1, 113-126.
  • Abel, S.D.A., & Barid, S.K. (2018). Honey is cytotoxic towards prostate cancer cells but interacts with the MTT reagent: Considerations for the choice of cell viability assay. Food Chemistry, 241, 70-78. https://doi: 10.1016/j.foodchem.2017.08.083
  • Bergman, J. 1998. The structure and properties of some indolic constituents in Couroupita guianensis aubl. Tetrahedron, 41(14), 2879. https://doi.org/10.1016/S0040-4020(01)96609-8
  • Cao, L., Zhang, L., Zhao, X., & Zhang, Y. (2016). A hybrid chalcone combining the trimethoxyphenyl and isatinyl groups targets multiple oncogenic proteins and pathways in hepatocellular carcinoma cells. PLoS ONE, 11(8), e0161025. https://doi:10.1371/journal.pone.0161025
  • Chiyanzu, I. (2003). Synthesis and evaluation of isatins and thiosemicarbazone derivatives against cruzain, falcipain-2 and rhodesain. Bioorganic & Medicinal Chemistry Letters, 13(20), 3527–3530. https://doi:10.1016/s0960-894x(03)00756-x
  • Chuang, S.C., Vecchia, C.L., & Boffetta, P. (2009). Liver cancer: descriptive epidemiology and risk factors other than HBV and HCV infection. Cancer Letters, 286(1), 9–14. https://doi:10.1016/j.canlet.2008.10.040
  • Eldeeb, M., Sanad, E.F., Ragab, A., Ammar, Y.A., Mahmoud, K., Ali, M.M., & Hamdy, N.M. (2022). Anticancer effects with molecular docking confirmation of newly synthesized isatin sulfonamide molecular hybrid derivatives against hepatic cancer cell lines. Biomedicines, 10(3), 722. https://doi: 10.3390/biomedicines10030722
  • Ferlay, J., Ervik, M., Lam, F., Colombet, M., Mery, L., & Piñeros, M. (2021). Global cancer observatory: cancer today. Lyon, France: International Agency for Research on Cancer. https://doi:10.1002/ijc.33588
  • Hossain, W., Hossain, M., Arafath, K., Ety, S.S., Shetu, M.H., Kabir, M., Noor, F.A., & Manoor, K. (2022). Real-Time fast PCR amplification using designated and conventional real time thermal cycler systems: COVID-19 perspective. PLOS ONE, 17(10), e0276464. https://doi:10.1371/journal.pone.0276464
  • Jaksch, M., Munera, J., Bajpai, R., Terskikh, A., & Oshima R.G. (2008). Cell cycle-dependent variation of a CD133 epitope in human embryonic stem cell, colon cancer, and melanoma cell lines. Cancer Research, 68(19), 7882–7886. https://doi:10.1158/0008-5472.CAN-08-0723
  • Jiang, T., Kuhen, K.L., Wolff, K., Yin, H., Bieza, K., Caldwell, J., Bursulaya, B., Wu, T.Y., & He, Y. (2006). Design, synthesis, and biological evaluations of novel oxindoles as HIV-1 non-nucleoside reverse transcriptase inhibitors. Bioorganic & Medicinal Chemistry Letters, 16(8), 2109–2112. https://doi: 10.1016/j.bmcl.2006.01.073
  • Jonkman, J.E.N., Cathcart, J.A., Xu, F., Bartolini, M.E., Amon, J.E., Stevens, K.M., & Colarusso, P. (2014). An introduction to the wound healing assay using live-cell microscopy. Cell Adhesion & Migration, 8(5), 440-451. https://doi:10.4161/cam.36224
  • Mallamo, J.P. (2006). Structure-guided identification of novel VEGFR-2 kinase inhibitors via solution phase parallel synthesis. Bioorganic & Medicinal Chemistry Letters, 16(8), 2158–2162. https://doi:10.1016/j.bmcl.2006.01.063
  • Marrero, J.A., Kulik, L.M, & Sirlin, C.B. (2018). Diagnosis, staging, and management of hepatocellular carcinoma: 2018 practice guidance by the American association for the study of liver diseases. Hepatology, 68(2), 723–750. https://doi:10.1002/hep.29913
  • McGlynn, K.A., Petrick J.L., & El-Serag, H.B. (2021). Epidemiology of hepatocellular carcinoma. Hepatology, 73, 4–13. https://doi:10.1002/hep.31288
  • Nagarsenkar, A., Guntuku, L., Guggilapu, S.D., Danthi, B.K., Gannoju, S., Naidu, V.G.M., & Bathini, N.B. (2016). Synthesis and apoptosis inducing studies of triazole linked 3-benzylidene isatin derivatives. European Journal of Medicinal Chemistry, 124, 782–793. https://doi:10.1016/j.ejmech.2016.09.009
  • Noufal, K.P., Rajesh, B., & Sujith, S. N. (2023). Antioxidant and cytotoxic effects of the methanolic extract of eichhornia crassipes petioles upon mg-63 cell lines: an ın vitro study. Cureus, 15(5): e38425. https://doi:10.7759/cureus.38425
  • Pandeya, S.N., Smitha, S., Jyoti, M., & Sridhar S.K. (2005). Biological activities of isatin and its derivatives. Acta Pharmaceutica, 55, 27–46.
  • Rumgay, H., Arnold, M., Ferlay, J., Lesi, O., Cabasag, C.J., Vignat, J., Laversanne, M., McGlynn, K.A., & Soerjomataram, I. (2022). Global burden of primary liver cancer in 2020 and predictions to 2040. Journal of Hepatology, 77(6), 1598–1606. https://doi:10.1016/j.jhep.2022.08.021
  • Silva, J.F.M., Garden, S.J., &Pinto, A.C. (2001). The chemistry of isatins: a review from 1975 to 1999. Journal of Brazilian Chemical Society, 12(3), 273. https://doi.org/10.1590/S0103-50532001000300002
  • Sriram, D. (2005). Synthesis and evaluation of anti-HIV activity of isatin beta-thiosemicarbazone derivatives. Bioorganic & Medicinal Chemistry Letters, 15(20), 4451–4455. https://doi:10.1016/j.bmcl.2005.07.046
  • Sung, H., Ferlay, J.,Siegel, R.L., Laversanne, M., Soerjomataram, I., & Jemal, A. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 71(3), 209-249. https://doi:10.3322/caac.21660
  • Tawfik, N.G., Mohamed W.R., Mahmoud, H.S., Alqarni, M.A., Naguib, I.A., Fahmy, A.M., & Ahmed, O.M. (2022). Isatin counteracts diethylnitrosamine/2-acetylaminofluoreneinduced hepatocarcinogenesis in male wistar rats by upregulating anti-inflammatory, antioxidant, and detoxification pathways. Antioxidants, 11(4), 699. https://doi.org/10.3390/antiox11040699

Investigation of the molecular effects of isatin on HepG2 and AML12 cell lines

Year 2024, Volume: 14 Issue: 2, 394 - 401, 15.06.2024
https://doi.org/10.17714/gumusfenbil.1389743

Abstract

Isatin is an indole-derived organic compound. It is a natural component of the Couroupita guianensis plant. It is also the metabolic derivative of the human body hormone adrenaline. Studies have shown the anti-tumoral effects of isatin derivatives. In this study, the cytotoxic effects of isatin on HepG2, a hepatocellular cancer cell line was investigated. Additionally, its cytotoxic and protective-proliferative effects on AML12, a healthy liver cell line was investigated. This evaluation was conducted using MTT, fluorescent staining, wound healing, and real-time polymerase chain reaction analyses. The IC50 values for 48 hours of isatin application were calculated as 186.23 µM for HepG2 and 7.05 mM for AML12. The analysis of wound healing and fluorescent staining at varying doses of HepG2 application revealed suppression of proliferation and triggered apoptosis in HepG2 cells. In contrast, AML12 cells exhibited promoted proliferation under similar conditions. Moreover, the observed upregulation of oxidative stress genes CuZn/Mn-SOD and mitochondrial apoptotic pathway genes Bax, cleaveled-Cas3, APAF1, and p53 in HepG2 cells contrasted with their decreased expression in AML12 cell lines. These results suggest the potential of natural isatin as a promising anti-cancer agent for liver cancer cell lines and as a protective supplement for healthy liver cells.

References

  • Abdoul-Fadl, T., & Bin-Jubair, F.A.S. (2010). Anti-tubercular activity of isatin and derivatives. International Journal of Research in Pharmaceutical Sciences, 1, 113-126.
  • Abel, S.D.A., & Barid, S.K. (2018). Honey is cytotoxic towards prostate cancer cells but interacts with the MTT reagent: Considerations for the choice of cell viability assay. Food Chemistry, 241, 70-78. https://doi: 10.1016/j.foodchem.2017.08.083
  • Bergman, J. 1998. The structure and properties of some indolic constituents in Couroupita guianensis aubl. Tetrahedron, 41(14), 2879. https://doi.org/10.1016/S0040-4020(01)96609-8
  • Cao, L., Zhang, L., Zhao, X., & Zhang, Y. (2016). A hybrid chalcone combining the trimethoxyphenyl and isatinyl groups targets multiple oncogenic proteins and pathways in hepatocellular carcinoma cells. PLoS ONE, 11(8), e0161025. https://doi:10.1371/journal.pone.0161025
  • Chiyanzu, I. (2003). Synthesis and evaluation of isatins and thiosemicarbazone derivatives against cruzain, falcipain-2 and rhodesain. Bioorganic & Medicinal Chemistry Letters, 13(20), 3527–3530. https://doi:10.1016/s0960-894x(03)00756-x
  • Chuang, S.C., Vecchia, C.L., & Boffetta, P. (2009). Liver cancer: descriptive epidemiology and risk factors other than HBV and HCV infection. Cancer Letters, 286(1), 9–14. https://doi:10.1016/j.canlet.2008.10.040
  • Eldeeb, M., Sanad, E.F., Ragab, A., Ammar, Y.A., Mahmoud, K., Ali, M.M., & Hamdy, N.M. (2022). Anticancer effects with molecular docking confirmation of newly synthesized isatin sulfonamide molecular hybrid derivatives against hepatic cancer cell lines. Biomedicines, 10(3), 722. https://doi: 10.3390/biomedicines10030722
  • Ferlay, J., Ervik, M., Lam, F., Colombet, M., Mery, L., & Piñeros, M. (2021). Global cancer observatory: cancer today. Lyon, France: International Agency for Research on Cancer. https://doi:10.1002/ijc.33588
  • Hossain, W., Hossain, M., Arafath, K., Ety, S.S., Shetu, M.H., Kabir, M., Noor, F.A., & Manoor, K. (2022). Real-Time fast PCR amplification using designated and conventional real time thermal cycler systems: COVID-19 perspective. PLOS ONE, 17(10), e0276464. https://doi:10.1371/journal.pone.0276464
  • Jaksch, M., Munera, J., Bajpai, R., Terskikh, A., & Oshima R.G. (2008). Cell cycle-dependent variation of a CD133 epitope in human embryonic stem cell, colon cancer, and melanoma cell lines. Cancer Research, 68(19), 7882–7886. https://doi:10.1158/0008-5472.CAN-08-0723
  • Jiang, T., Kuhen, K.L., Wolff, K., Yin, H., Bieza, K., Caldwell, J., Bursulaya, B., Wu, T.Y., & He, Y. (2006). Design, synthesis, and biological evaluations of novel oxindoles as HIV-1 non-nucleoside reverse transcriptase inhibitors. Bioorganic & Medicinal Chemistry Letters, 16(8), 2109–2112. https://doi: 10.1016/j.bmcl.2006.01.073
  • Jonkman, J.E.N., Cathcart, J.A., Xu, F., Bartolini, M.E., Amon, J.E., Stevens, K.M., & Colarusso, P. (2014). An introduction to the wound healing assay using live-cell microscopy. Cell Adhesion & Migration, 8(5), 440-451. https://doi:10.4161/cam.36224
  • Mallamo, J.P. (2006). Structure-guided identification of novel VEGFR-2 kinase inhibitors via solution phase parallel synthesis. Bioorganic & Medicinal Chemistry Letters, 16(8), 2158–2162. https://doi:10.1016/j.bmcl.2006.01.063
  • Marrero, J.A., Kulik, L.M, & Sirlin, C.B. (2018). Diagnosis, staging, and management of hepatocellular carcinoma: 2018 practice guidance by the American association for the study of liver diseases. Hepatology, 68(2), 723–750. https://doi:10.1002/hep.29913
  • McGlynn, K.A., Petrick J.L., & El-Serag, H.B. (2021). Epidemiology of hepatocellular carcinoma. Hepatology, 73, 4–13. https://doi:10.1002/hep.31288
  • Nagarsenkar, A., Guntuku, L., Guggilapu, S.D., Danthi, B.K., Gannoju, S., Naidu, V.G.M., & Bathini, N.B. (2016). Synthesis and apoptosis inducing studies of triazole linked 3-benzylidene isatin derivatives. European Journal of Medicinal Chemistry, 124, 782–793. https://doi:10.1016/j.ejmech.2016.09.009
  • Noufal, K.P., Rajesh, B., & Sujith, S. N. (2023). Antioxidant and cytotoxic effects of the methanolic extract of eichhornia crassipes petioles upon mg-63 cell lines: an ın vitro study. Cureus, 15(5): e38425. https://doi:10.7759/cureus.38425
  • Pandeya, S.N., Smitha, S., Jyoti, M., & Sridhar S.K. (2005). Biological activities of isatin and its derivatives. Acta Pharmaceutica, 55, 27–46.
  • Rumgay, H., Arnold, M., Ferlay, J., Lesi, O., Cabasag, C.J., Vignat, J., Laversanne, M., McGlynn, K.A., & Soerjomataram, I. (2022). Global burden of primary liver cancer in 2020 and predictions to 2040. Journal of Hepatology, 77(6), 1598–1606. https://doi:10.1016/j.jhep.2022.08.021
  • Silva, J.F.M., Garden, S.J., &Pinto, A.C. (2001). The chemistry of isatins: a review from 1975 to 1999. Journal of Brazilian Chemical Society, 12(3), 273. https://doi.org/10.1590/S0103-50532001000300002
  • Sriram, D. (2005). Synthesis and evaluation of anti-HIV activity of isatin beta-thiosemicarbazone derivatives. Bioorganic & Medicinal Chemistry Letters, 15(20), 4451–4455. https://doi:10.1016/j.bmcl.2005.07.046
  • Sung, H., Ferlay, J.,Siegel, R.L., Laversanne, M., Soerjomataram, I., & Jemal, A. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 71(3), 209-249. https://doi:10.3322/caac.21660
  • Tawfik, N.G., Mohamed W.R., Mahmoud, H.S., Alqarni, M.A., Naguib, I.A., Fahmy, A.M., & Ahmed, O.M. (2022). Isatin counteracts diethylnitrosamine/2-acetylaminofluoreneinduced hepatocarcinogenesis in male wistar rats by upregulating anti-inflammatory, antioxidant, and detoxification pathways. Antioxidants, 11(4), 699. https://doi.org/10.3390/antiox11040699
There are 23 citations in total.

Details

Primary Language English
Subjects Cancer Biology
Journal Section Articles
Authors

Deniz Şumnulu 0009-0009-0693-3569

Publication Date June 15, 2024
Submission Date November 12, 2023
Acceptance Date January 9, 2024
Published in Issue Year 2024 Volume: 14 Issue: 2

Cite

APA Şumnulu, D. (2024). Investigation of the molecular effects of isatin on HepG2 and AML12 cell lines. Gümüşhane Üniversitesi Fen Bilimleri Dergisi, 14(2), 394-401. https://doi.org/10.17714/gumusfenbil.1389743