Araştırma Makalesi
Yıl 2020,
Cilt: 45 Sayı: 3, 1165 - 1172, 30.09.2020
Öz
Amaç: Çalışmamızda Hibiscus vitifolius’dan izole edilen ve antioksidan, antienflamatuar, analjezik ve anti kanser özelliklere sahip olan gossypinin Hep-3B hücreleri üzerindeki antiproliferatif etkilerini göstermeyi amaçladık.
Gereç ve Yöntem: Çalışmamızda Hep-3B hücre hatları American Type Culture Collection (ATCC, USA) temin edilmiştir. Hücreler farklı konsantrasyonlarda (5-100 µg/ml) gossypin ve pozitif kontrol olarak da sisplatine (50µM) maruz bırakılmışlardır. Sonrasında 24, 48 ve 72 saatlerde hücrelere (MTT) yöntemi ile canlılık analizi yapılmıştır. Hücrelerdeki apoptozun göstergesi için Hoechst floresan boyama yapıldı. Aynı zamanda RT-PCR ile Nuklear kappa B (NFκB), kaspaz 3 ve 9 mRNA ekspresyon düzeyleri incelendi.
Bulgular: Çalışmamızda gossypin uygulaması doza ve zamana bağlı olarak hücre proliferasyonunu önlemiştir. İlk 24 saatte sadece 100µg/ml dozunda etki gösterirken 48 ve 72. saatlerde ise doza bağlı olarak etkisini göstermiştir. Hoechst floresan boyama ile gossypin 50 ve 100 µg/ml dozunda hücreleri daha belirgin apoptoza götürdüğü görülürken sisplatin grubu ile neredeyse aynı etkiyi ortaya koymuştur. NFκB mRNA ekspresyonunu doza bağlı inhibe eden gossypin, aynı zamanda apoptotik protein olan kaspas 3 ve 9’un mRNA ekspresyonlarını doza bağlı olarak indüklemiştir.
Sonuç: Gossypin doza bağlı olarak Hep-3B hücreleri üzerinde sisplatin ile yakın bir etki ortaya koymuştur. Bu etkilerini ise apoptozu aktive ederek ve NFκB inhibisyonu yaparak ortaya koymuştur. Buradan yola çıkarak karaciğer kanserinin tedavisinde gossypinin gelecekte potansiyel bir antikanser ajan olabilir.
Destekleyen Kurum
Yok
Proje Numarası
Yok
Teşekkür
Yok
Kaynakça
- Referans1. London WT, Petrick JL, KA. M. Liver cancer. In: Thun MJ LM, Cerhan JR, Haiman CA, Schottenfeld D, editor. In Cancer Epidemiology and Prevention. 4th ed. New York: Oxford University Press; 2018. p. 635-60. Referans2. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364-78. Referans3. Ju SM, Kang JG, Bae JS, Pae HO, Lyu YS, Jeon BH. The Flavonoid Apigenin Ameliorates Cisplatin-Induced Nephrotoxicity through Reduction of p53 Activation and Promotion of PI3K/Akt Pathway in Human Renal Proximal Tubular Epithelial Cells. Evid Based Complement Alternat Med. 2015;2015:186436. Referans4. Lavery HJ, Cooperberg MR. Clinically localized prostate cancer in 2017: A review of comparative effectiveness. Urol Oncol. 2017;35:40-1. Referans5. Vijayan P, Raghu C, Ashok G, Dhanaraj SA, Suresh B. Antiviral activity of medicinal plants of Nilgiris. Indian J Med Res. 2004;120:24-9. Referans6. Gautam P, Flora SJ. Oral supplementation of gossypin during lead exposure protects alteration in heme synthesis pathway and brain oxidative stress in rats. Nutrition. 2010;26:563-70. Referans7. Venkatesan T, Sorimuthu Pillai S. Antidiabetic activity of gossypin, a pentahydroxyflavone glucoside, in streptozotocin-induced experimental diabetes in rats. J Diabetes. 2012;4:41-6. Referans8. Viswanathan S, Sambantham PT, Reddy K, Kameswaran L. Gossypin-induced analgesia in mice. Eur J Pharmacol. 1984;98:289-91. Referans9. Yoon I, Lee KH, Cho J. Gossypin protects primary cultured rat cortical cells from oxidative stress- and beta-amyloid-induced toxicity. Arch Pharm Res. 2004;27:454-9. Referans10. Ramaswamy S, Viswanathan S. Influence of gossypin on the development of acute tolerance to morphine induced antinociception. Indian J Exp Biol. 1997;35:413-4. Referans11. Anon MT, Ubeda A, Alcaraz MJ. Protective effects of phenolic compounds on CCl4-induced toxicity in isolated rat hepatocytes. Z Naturforsch C J Biosci. 1992;47:275-9. Referans12. Cinar I, Sirin B, Aydin P, Toktay E, Cadirci E, Halici I, et al. Ameliorative effect of gossypin against acute lung injury in experimental sepsis model of rats. Life Sci. 2019;221:327-34. Referans13. Popperl G, Helmberger T, Munzing W, Schmid R, Jacobs TF, Tatsch K. Selective internal radiation therapy with SIR-Spheres in patients with nonresectable liver tumors. Cancer Biother Radiopharm. 2005;20:200-8. Referans14. Shaghayegh K, A Mahdi, K Ali. Larynx Preserving Treatments in the Early and Advanced Laryngeal Cancers; a Retrospective Analysis. J Cancer Sci Ther. 2010;1:8-10. Referans15. Babu BH, Jayram HN, Nair MG, Ajaikumar KB, Padikkala J. Free radical scavenging, antitumor and anticarcinogenic activity of gossypin. J Exp Clin Cancer Res. 2003;22:581-9. Referans16. Shi L, Chen J, Wang YY, Sun G, Liu JN, Zhang JX, et al. Gossypin induces G2/M arrest in human malignant glioma U251 cells by the activation of Chk1/Cdc25C pathway. Cell Mol Neurobiol. 2012;32:289-96. Referans17. Wang L, Wang X, Chen H, Zu X, Ma F, Liu K, et al. Gossypin inhibits gastric cancer growth by direct targeting of AURKA and RSK2. Phytother Res. 2019;33:640-50. Referans18. van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011;731:237-45. Referans19. Plumb JA. Cell sensitivity assays : the MTT assay. Methods Mol Med. 1999;28:25-30. Referans20. Nikkhah G, Tonn JC, Hoffmann O, Kraemer HP, Darling JL, Schachenmayr W, et al. The MTT assay for chemosensitivity testing of human tumors of the central nervous system. Part II: Evaluation of patient- and drug-specific variables. J Neurooncol. 1992;13:13-24. Referans21. Yeh TC, Chiang PC, Li TK, Hsu JL, Lin CJ, Wang SW, et al. Genistein induces apoptosis in human hepatocellular carcinomas via interaction of endoplasmic reticulum stress and mitochondrial insult. Biochem Pharmacol. 2007;73:782-92. Referans22. Shi Y, Zhang L, Liu X, Zhou C, Zhang L, Zhang S, et al. Icotinib versus gefitinib in previously treated advanced non-small-cell lung cancer (ICOGEN): a randomised, double-blind phase 3 non-inferiority trial. Lancet Oncol. 2013;14:953-61. Referans23. Vaux DL, Haecker G, Strasser A. An evolutionary perspective on apoptosis. Cell. 1994;76:777-9. Referans24. Horvitz HR. Genetic control of programmed cell death in the nematode Caenorhabditis elegans. Cancer Res. 1999;59:1701s-6s. Referans25. Wong RS. Apoptosis in cancer: from pathogenesis to treatment. J Exp Clin Cancer Res. 2011;30:87. Referans26. Kroemer G, Galluzzi L, Brenner C. Mitochondrial membrane permeabilization in cell death. Physiol Rev. 2007;87:99-163. Referans27. Tsujimoto Y, Finger LR, Yunis J, Nowell PC, Croce CM. Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science. 1984;226:1097-9. Referans28. Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science. 2004;305:626-9. Referans29. Decaudin D, Marzo I, Brenner C, Kroemer G. Mitochondria in chemotherapy-induced apoptosis: a prospective novel target of cancer therapy (review). Int J Oncol. 1998;12:141-52. Referans30. Kunnumakkara AB, Nair AS, Ahn KS, Pandey MK, Yi Z, Liu M, et al. Gossypin, a pentahydroxy glucosyl flavone, inhibits the transforming growth factor beta-activated kinase-1-mediated NF-kappaB activation pathway, leading to potentiation of apoptosis, suppression of invasion, and abrogation of osteoclastogenesis. Blood. 2007;109:5112-21. Referans31. Zhivotosky B, Orrenius S. Assessment of apoptosis and necrosis by DNA fragmentation and morphological criteria. Curr Protoc Cell Biol. 2001;Chapter 18:18 3 1- 3 23. Referans32. Errami Y, Naura AS, Kim H, Ju J, Suzuki Y, El-Bahrawy AH, et al. Apoptotic DNA fragmentation may be a cooperative activity between caspase-activated deoxyribonuclease and the poly(ADP-ribose) polymerase-regulated DNAS1L3, an endoplasmic reticulum-localized endonuclease that translocates to the nucleus during apoptosis. J Biol Chem. 2013;288:3460-8. Referans33. Matassov D, Kagan T, Leblanc J, Sikorska M, Zakeri Z. Measurement of apoptosis by DNA fragmentation. Methods Mol Biol. 2004;282:1-17. Referans34. Hui F, Qin X, Zhang Q, Li R, Liu M, Ren T, et al. Alpinia oxyphylla oil induces apoptosis of hepatocellular carcinoma cells via PI3K/Akt pathway in vitro and in vivo. Biomed Pharmacother. 2019;109:2365-74.
Yıl 2020,
Cilt: 45 Sayı: 3, 1165 - 1172, 30.09.2020
Öz
Purpose: The aim of this study was to demonstrate the antiproliferative effects of Gossypin which has antioxidant, anti-inflammatory, analgesic and anti-cancer properties on Hep-3B cells isolated from Hibiscus vitifolius.
Materials and Methods: Hep-3B Cell Lines American Type Culture Collection (ATCC, USA) was provided in our study. The cells were exposed to different concentrations (5-100 µg/ml) of gossypin and cisplatine (50µM) as positive control. Afterwards, viability analysis was performed with MTT method on the cells at 24, 48 and 72 hours. Hoechst fluorescent staining was performed for indication of apoptosis in cells. At the same time, RT-PCR and NFκB, caspase 3 and 9 mRNA expression levels were examined.
Results: Gossypin administration prevented cell proliferation due to dose and time. In the first 24 hours, only 100µg/ml dose was effective, while in 48 and 72 hours it was effective depending on the dose. With Hoechst fluorescent staining, gossypin showed nearly the same effect as the cisplatin group, with doses of 50 and 100 µg/ml seen to lead the cells to more apoptosis. Gossypin, which inhibits dose dependent NFκB mRNA expression, has also up regulated dose dependent the apoptotic proteins mRNA expression of caspase 3 and 9.
Conclusion: Gossypin showed a close effect with cisplatin on Hep-3B cells depending on the dose. These effects were demonstrated by activating apoptosis and inhibiting NFκB. Based on this, gossypin may be a potential anticancer agent in the future for the treatment of liver cancer.
Proje Numarası
Yok
Kaynakça
- Referans1. London WT, Petrick JL, KA. M. Liver cancer. In: Thun MJ LM, Cerhan JR, Haiman CA, Schottenfeld D, editor. In Cancer Epidemiology and Prevention. 4th ed. New York: Oxford University Press; 2018. p. 635-60. Referans2. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364-78. Referans3. Ju SM, Kang JG, Bae JS, Pae HO, Lyu YS, Jeon BH. The Flavonoid Apigenin Ameliorates Cisplatin-Induced Nephrotoxicity through Reduction of p53 Activation and Promotion of PI3K/Akt Pathway in Human Renal Proximal Tubular Epithelial Cells. Evid Based Complement Alternat Med. 2015;2015:186436. Referans4. Lavery HJ, Cooperberg MR. Clinically localized prostate cancer in 2017: A review of comparative effectiveness. Urol Oncol. 2017;35:40-1. Referans5. Vijayan P, Raghu C, Ashok G, Dhanaraj SA, Suresh B. Antiviral activity of medicinal plants of Nilgiris. Indian J Med Res. 2004;120:24-9. Referans6. Gautam P, Flora SJ. Oral supplementation of gossypin during lead exposure protects alteration in heme synthesis pathway and brain oxidative stress in rats. Nutrition. 2010;26:563-70. Referans7. Venkatesan T, Sorimuthu Pillai S. Antidiabetic activity of gossypin, a pentahydroxyflavone glucoside, in streptozotocin-induced experimental diabetes in rats. J Diabetes. 2012;4:41-6. Referans8. Viswanathan S, Sambantham PT, Reddy K, Kameswaran L. Gossypin-induced analgesia in mice. Eur J Pharmacol. 1984;98:289-91. Referans9. Yoon I, Lee KH, Cho J. Gossypin protects primary cultured rat cortical cells from oxidative stress- and beta-amyloid-induced toxicity. Arch Pharm Res. 2004;27:454-9. Referans10. Ramaswamy S, Viswanathan S. Influence of gossypin on the development of acute tolerance to morphine induced antinociception. Indian J Exp Biol. 1997;35:413-4. Referans11. Anon MT, Ubeda A, Alcaraz MJ. Protective effects of phenolic compounds on CCl4-induced toxicity in isolated rat hepatocytes. Z Naturforsch C J Biosci. 1992;47:275-9. Referans12. Cinar I, Sirin B, Aydin P, Toktay E, Cadirci E, Halici I, et al. Ameliorative effect of gossypin against acute lung injury in experimental sepsis model of rats. Life Sci. 2019;221:327-34. Referans13. Popperl G, Helmberger T, Munzing W, Schmid R, Jacobs TF, Tatsch K. Selective internal radiation therapy with SIR-Spheres in patients with nonresectable liver tumors. Cancer Biother Radiopharm. 2005;20:200-8. Referans14. Shaghayegh K, A Mahdi, K Ali. Larynx Preserving Treatments in the Early and Advanced Laryngeal Cancers; a Retrospective Analysis. J Cancer Sci Ther. 2010;1:8-10. Referans15. Babu BH, Jayram HN, Nair MG, Ajaikumar KB, Padikkala J. Free radical scavenging, antitumor and anticarcinogenic activity of gossypin. J Exp Clin Cancer Res. 2003;22:581-9. Referans16. Shi L, Chen J, Wang YY, Sun G, Liu JN, Zhang JX, et al. Gossypin induces G2/M arrest in human malignant glioma U251 cells by the activation of Chk1/Cdc25C pathway. Cell Mol Neurobiol. 2012;32:289-96. Referans17. Wang L, Wang X, Chen H, Zu X, Ma F, Liu K, et al. Gossypin inhibits gastric cancer growth by direct targeting of AURKA and RSK2. Phytother Res. 2019;33:640-50. Referans18. van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011;731:237-45. Referans19. Plumb JA. Cell sensitivity assays : the MTT assay. Methods Mol Med. 1999;28:25-30. Referans20. Nikkhah G, Tonn JC, Hoffmann O, Kraemer HP, Darling JL, Schachenmayr W, et al. The MTT assay for chemosensitivity testing of human tumors of the central nervous system. Part II: Evaluation of patient- and drug-specific variables. J Neurooncol. 1992;13:13-24. Referans21. Yeh TC, Chiang PC, Li TK, Hsu JL, Lin CJ, Wang SW, et al. Genistein induces apoptosis in human hepatocellular carcinomas via interaction of endoplasmic reticulum stress and mitochondrial insult. Biochem Pharmacol. 2007;73:782-92. Referans22. Shi Y, Zhang L, Liu X, Zhou C, Zhang L, Zhang S, et al. Icotinib versus gefitinib in previously treated advanced non-small-cell lung cancer (ICOGEN): a randomised, double-blind phase 3 non-inferiority trial. Lancet Oncol. 2013;14:953-61. Referans23. Vaux DL, Haecker G, Strasser A. An evolutionary perspective on apoptosis. Cell. 1994;76:777-9. Referans24. Horvitz HR. Genetic control of programmed cell death in the nematode Caenorhabditis elegans. Cancer Res. 1999;59:1701s-6s. Referans25. Wong RS. Apoptosis in cancer: from pathogenesis to treatment. J Exp Clin Cancer Res. 2011;30:87. Referans26. Kroemer G, Galluzzi L, Brenner C. Mitochondrial membrane permeabilization in cell death. Physiol Rev. 2007;87:99-163. Referans27. Tsujimoto Y, Finger LR, Yunis J, Nowell PC, Croce CM. Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science. 1984;226:1097-9. Referans28. Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science. 2004;305:626-9. Referans29. Decaudin D, Marzo I, Brenner C, Kroemer G. Mitochondria in chemotherapy-induced apoptosis: a prospective novel target of cancer therapy (review). Int J Oncol. 1998;12:141-52. Referans30. Kunnumakkara AB, Nair AS, Ahn KS, Pandey MK, Yi Z, Liu M, et al. Gossypin, a pentahydroxy glucosyl flavone, inhibits the transforming growth factor beta-activated kinase-1-mediated NF-kappaB activation pathway, leading to potentiation of apoptosis, suppression of invasion, and abrogation of osteoclastogenesis. Blood. 2007;109:5112-21. Referans31. Zhivotosky B, Orrenius S. Assessment of apoptosis and necrosis by DNA fragmentation and morphological criteria. Curr Protoc Cell Biol. 2001;Chapter 18:18 3 1- 3 23. Referans32. Errami Y, Naura AS, Kim H, Ju J, Suzuki Y, El-Bahrawy AH, et al. Apoptotic DNA fragmentation may be a cooperative activity between caspase-activated deoxyribonuclease and the poly(ADP-ribose) polymerase-regulated DNAS1L3, an endoplasmic reticulum-localized endonuclease that translocates to the nucleus during apoptosis. J Biol Chem. 2013;288:3460-8. Referans33. Matassov D, Kagan T, Leblanc J, Sikorska M, Zakeri Z. Measurement of apoptosis by DNA fragmentation. Methods Mol Biol. 2004;282:1-17. Referans34. Hui F, Qin X, Zhang Q, Li R, Liu M, Ren T, et al. Alpinia oxyphylla oil induces apoptosis of hepatocellular carcinoma cells via PI3K/Akt pathway in vitro and in vivo. Biomed Pharmacother. 2019;109:2365-74.
Toplam 1 adet kaynakça vardır.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Klinik Tıp Bilimleri (Diğer) |
| Bölüm | Araştırma |
| Yazarlar | |
| Proje Numarası | Yok |
| Yayımlanma Tarihi | 30 Eylül 2020 |
| Kabul Tarihi | 1 Temmuz 2020 |
| Yayımlandığı Sayı | Yıl 2020 Cilt: 45 Sayı: 3 |
Cited By
