Nor-wogonin’in İnsan Rahim Ağzı Kanseri Hücrelerinde Antikanser ve Apoptotik Etkileri

Yıl 2022, Sayı: 34, 617 - 622, 31.03.2022

Ahmet Karakuş , Sevgi Ünal Karakuş

https://doi.org/10.31590/ejosat.1083755

Cited By: 1

Öz

Tüm dünyada büyüyen bir sağlık sorunu olan kanser, kardiyovasküler hastalıklardan sonra ikinci en önemli ölüm nedenidir. Kanser tedavisinde kullanılan klasik yöntemlerin (cerrahi, radyoterapi, kemoterapi vb.) tedavide istenilen düzeyde etkili olamaması araştırmacıları kanser tedavisinde yeni terapötiklerin keşfine ve geliştirilmesine yönlendirmiştir. Flavonoidlerin bir alt grubu olan flavonların biyokimyasal ve farmakolojik etkilerine ek olarak antikanser, antienflamatuvar, antiproleratif, antianjiyogenik etkileri olduğu bilinmektedir. Çeşitli biyolojik aktivitelere sahip bir polihidroksi flavon türevi olan Nor-wogonin'in (5, 7, 8-trihydroxyflavone) insan rahim ağzı kanseri hücre hattı üzerine sitotoksik etkisini ve apoptoza dahil olan bazı genlerin eksprosyon düzeyine etkisini belirlemeyi amaçladık. Çalışmada, rahim ağzı (HeLa) kanser hücre hattına farklı konsantrasyonlarda (10, 20, 40, 80 µM) Nor-wogonin uygulandı. Bileşiğin hücre hatları üzerindeki sitotoksik etkisi MTT (3-(4,5-dimetiltiyazol-2-il)-difenil tetrazolyum bromür) yöntemi ile belirlendi ve HeLa hücreleri için Nor-wogonin IC50 değeri hesaplandı. Ek olarak, Nor-wogonin'in HeLa hücrelerindeki anti-apoptotik ve pro-apoptotik gen ekspresyon (bax, p53, caspase-3 ve bcl-2 genleri) seviyeleri üzerindeki etkisini gözlemlemek için Q-PCR ile kantitatif gen analizi yapıldı. Nor-wogonin'in artan doza bağlı olarak HeLa hücrelerinde hücre canlılığını önemli ölçüde azalttığı belirlendi (p<0.05). HeLa hücreleri için IC50 değerleri 32.09 µM olarak hesaplandı. Ayrıca HeLa hücrelerinde bax, kaspaz-3, p53 gen ekspresyonlarının arttığı ve bcl-2 gen ekspresyonunun azaldığı belirlendi. Sonuç olarak, bir flavon türevi olan Nor-wogonin, in vitro sitotoksik aktivitesi ile anti-apoptotik ve pro-apoptotik gen ekspresyonu üzerindeki etkileri sayesinde önemli bir terapötik ajan olarak kabul edilebilir.

Anahtar Kelimeler

Nor-wogonin, Rahim ağzı kanseri, Antikanser etki, Apoptotik Etki, Hücre kültürü.

Destekleyen Kurum

Bartın Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi

Proje Numarası

2021-FEN-B-001

Teşekkür

Desteklerinden dolayı Bartın Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi’ne teşekkür ederiz.

Anticancer and Apoptotic Effects of Nor-wogonin on Human Cervical Cancer Cells

Öz

Cancer, a growing health problem all over the world, is the second most important cause of death after cardiovascular diseases. The ineffectiveness of classical methods (surgery, radiotherapy, chemotherapy, etc.) used in cancer treatment has led researchers to discover and develop new therapeutics in cancer treatment. It is known that flavones, a subgroup of flavonoids, have anticancer, anti-inflammatory, antiprolerative, and antiangiogenic effects in addition to their biochemical and pharmacological effects. We aimed to determine the cytotoxic effect of Nor-wogonin (5, 7, 8-trihydroxyflavone), a polyhydroxyflavone derivative with various biological activities, on the human cervical cancer cell line and its effect on the expression level of some genes involved in apoptosis. In the study, different concentrations of Nor-wogonin (10, 20, 40, 80 µM) were applied to the cervical (HeLa) cancer cell line. The cytotoxic effect of the compound on cell lines was determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-diphenyl tetrazolium bromide) method and the Nor-wogonin IC50 value for HeLa cells was calculated. In addition, quantitative gene analysis by Q-PCR was performed to observe the effect of Nor-wogonin on the levels of anti-apoptotic and pro-apoptotic gene expression (bax, p53, caspase-3 and bcl-2 genes) in HeLa cells. It was determined that Nor-wogonin significantly decreased cell viability in HeLa cells depending on the increasing dose (p<0.05). IC50 values for HeLa cells were calculated as 32.09 µM. In addition, it was determined that bax, caspase-3, p53 gene expressions increased and bcl-2 gene expression decreased in HeLa cells. In conclusion, Nor-wogonin, a flavone derivative, can be considered as an important therapeutic agent thanks to its in vitro cytotoxic activity and its effects on anti-apoptotic and pro-apoptotic gene expression.

Anahtar Kelimeler

Nor-wogonin, Cervical cancer, Antikanser effect, Apoptotic effect, Cell culture.

Proje Numarası

2021-FEN-B-001

Kaynakça

• Abd El-Hafeez, A. A., Khalifa, H. O., Mahdy, E. A. M., Sharma, V., Hosoi T., Ghosh, P., et al. (2019). Anticancer effect of nor-wogonin (5, 7, 8-trihydroxyflavone) on human triple-negative breast cancer cells via downregulation of TAK1, NF-κB, and STAT3. Pharmacol Rep. 71(2), 289-298.
• Banik, K., Khatoon, E., Harsha, C., Rana, V., Parama, D., Thakur, K. K., Bishayee, A., Kunnumakkara, A. B. (2022). Wogonin and its analogs for the prevention and treatment of cancer: A systematic review. Phytotherapy Research.
• Bose, S., Banerjee, S., Mondal, A., Chakraborty, U., Pumarol, J., Croley, C. R., & Bishayee, A. (2020). Targeting the JAK/STAT signaling pathway using Phytocompounds for cancer prevention and therapy. Cell, 9(6), 1451.
• Ceylan, H., Budak, H., Kocpinar, E. F., Baltaci, N. G. & Erdogan, O. (2019). Examining the link between dose-dependent dietary iron intake and Alzheimer’s disease through oxidative stress in the rat cortex. J Trace Elem Med Biol. 56, 198-206.
• Chipuk, J. E., Kuwana, T., Bouchier-Hayes, L., Droin, N. M., Newmeyer, D. D., Schuler, M., et al. (2004). Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science, 303, 1010-1014.
• Chow, J. M., Huang, G. C., Shen, S. C., Wu, C. Y., Lin, C. W. & Chen, Y. C. (2008). Differential apoptotic effect of wogonin and nor-wogonin via stimulation of ROS production in human leukemia cells. J Cell Biochem,103, 1394–1404.
• Cragg, G. M., & Pezzuto, J. M. (2016). Natural products as a vital source for the discovery of cancer chemotherapeutic and Chemopreventive agents. Medical Principles and Practice, 25, 41–59
• Daimary, U. D., Dey, P., Rana, V., Banik, K., Kumar, A., Choudhary, H., & Kunnumakkara, A. (2020). Emerging roles of cardamonin, a multitargeted nutraceutical in the prevention and treatment of chronic diseases. Curr Res Pharmacol Drug Discov, 10, 2:100008.
• Harsha, C., Banik, K., Bordoloi, D., & Kunnumakkara, A. B. (2017). Antiulcer properties of fruits and vegetables: A mechanism based perspective. Food and Chemical Toxicology, 108(Pt A), 104–119.
• Havsteen, B. H. (2002). The biochemistry and medical significance of the flavonoids. Pharmacology and Therapeutics, 96, 67-202.
• Hui, K. M, Huen, M. S., Wang, H. Y., Zheng, H., Sigel, E., Baur, R., et al. (2002). Anxiolytic effect of wogonin, a benzodiazepine receptor ligand isolated from Scutellaria baicalensis Georgi. Biochem Pharmacol, 64(9), 1415-1424.
• Huynh, D. L., Sharma, N., Singh, A. K., Sodhi, S. S., Zhang, J. J., Mongre, R. K., Ghosh, M., Kim, N., Park, Y. H. & Jeong, D. K. (2017). Anti-tumor activity of wogonin, an extract from Scutellaria baicalensis, through regulating different signaling pathways. Chin J Nat Med, 15(1), 15–40.
• Khatoon, E., Banik, K., Harsha, C., Sailo, B. L., Thakur, K. K., Khwairakpam, A. D., et al. (2020). Phytochemicals in cancer cell chemosensitization: Current knowledge and future perspectives. Seminars in Cancer Biology, S1044-579X(20), 30150–4.
• Koran, K., et al., (2017). Synthesis, structural and thermal characterizations and in vitro cytotoxic activities of new cyclotriphosphazene derivatives. Phosphorus, Sulfur, and Silicon and the Related Elements, 192(9), 1002-1011.
• Liu, J., Huang, R., Lin, D., Peng, J., Wu, X. & Lin, Q. (2005). Expression of survivin and bax/bcl-2 in peroxisome proliferator activated receptor-γ ligands induces apoptosis on human myeloid leukemia cells in vitro. Ann Oncol,16, 455-459.
• Livak, K. J. & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25(4), 402-408.
• Martens, S. & Mithöfer, A. (2205). Flavones and flavone synthases. Phytochemistry, 66(20), 2399-2407.
• Miyasaki, Y., Rabenstein, J. D., Rhea, J., Crouch, M. L., Mocek, U. M., Kittell, P. E., et al. (2013). Isolation and characterization of antimicrobial compounds in plant extracts against multidrug. PlosOne,8, 4.
• Miyashita, T., Krajewski, S., Krajewska, M., Wang, H. G., Lin, H., Liebermann, D. A., et al. (1994). Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene, 9, 1799-1805.
• Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983. 65(1), 55-63.
• Parama, D., Boruah, M., Yachna, K., Rana, V., Banik, K., Harsha, C., et al. (2020). Diosgenin, a steroidal saponin, and its analogs: Effective therapies against different chronic diseases. Life Sciences, 260, 118182.
• Ravishankar, D., Rajora, A. K, Greco, F. & Osborn, H. M. I. (2013). Flavonoids as prospective compounds for anti-cancer therapy. Int J Biochem Cell Biol, 45, 2821-2831.
• S Herzog, T. J & Wright J. D. (2007). The impact of cervical cancer on quality of life--the components and means for management. Gynecol Oncol, 107(3), 572-577.
• Singh, M., Maninder, K. & Om, Si. (2014). Flavones: An important scaffold for medicinal chemistry. Eur J Med Chem, 84, 206-239.
• Srivastava, S., Somasagara, R. R., Hegde, M., Nishana, M., Tadi, S. K, Srivastava, M, et al. (2016). Natural Flavonoid Interacts with DNA, Arrests Cell Cycle and Causes Tumor Regression by Activating Mitochondrial Pathway of Apoptosis. Sci Rep, (12)6, 24049.
• Tewari, D., Patni, P., Bishayee, A., Sah, A. N., & Bishayee, A. (2019). Natural products targeting the PI3K-Akt-mTOR signaling pathway in cancer: A novel therapeutic strategy. Seminars in Cancer Biology, (19), 30405–5.
• Waggoner, S. E. (2003). Cervical cancer. The Lancet, (361)9376, 2217-2225.
• Wang, Z., Zhang, Q., Zhou, L., Liu, G., Wu, Q. & Chen, C. (2020). Nor-wogonin flavone suppresses the growth of human colon cancer cells via mitochondrial mediated apoptosis, autophagy induction and triggering G2/M phase cell cycle arrest. J Buon, 25(3), 1449-1454.