RUTİNİN EPİTELYAL VE MEZENKİMAL GEÇİŞ ÜZERİNE OLAN ETKİSİNİN PROSTAT KANSERİ HÜCRELERİNDE BELİRLENMESİ
Öz
Amaç: Prostat kanseri tedavisinde yaşanan zorluklar, kemoterapi nedeniyle görülen yan etkilerin fazlalığı alternatif tedavi stratejileri arayışını gündeme getirmiştir. Son yıllarda yapılan araştırmalarda Rutin (RUT)’in kanser hücreleri üzerinde anti-kanser etki gösterdiği bilinmektedir. Bu nedenle çalışmamızda RUT’un prostat kanseri hücrelerinde epitelyal mezenkimal geçiş (EMT) üzerindeki etkilerinin ilk defa belirlenmesi amaçlanmıştır.
Yöntem: Prostat kanseri hücrelerinde (PC-3) RUT’un antikanser aktivitesi WST-1, Annexin V ELISA, DAPI boyama ve Akridin Oranj (AO) boyama ile belirlenerek, RUT’un anti-kanser ve anti-metastatik özellikleri Scratch (Çizik) Assay testi ile değerlendirildi. Bax, Bcl-2, Snail, Twist, Vimentin ve E-kaderin genlerinin mRNA ifade düzeyi RT-PCR analizi ile belirlendi.
Bulgular: PC-3 hücrelerine, farklı konstrasyonlarda (500, 750, 1000, 1500 µM) ve farklı zamanlarda (24 ve 48 saat) RUT uygulandı. Canlılık oranlarının doza ve zamana bağlı olarak artan RUT konsantrasyonuna bağlı olarak azaldığı gözlendi (p<0,01). Ayrıca artan RUT konsantrasyonuna bağlı olarak Annexin V proteinin azaldığı ve apoptotik hücrelerin arttığı belirlendi (p<0,01). Özellikle 750 ve 1000 µM RUT konsantrasyonlarında göç eden hücre sayısında azalma olduğu ortaya kondu. RT-PCR analizi sonucu elde edilen verilerde 1500 µM RUT muamelesi sonrası E-Kaderin mRNA seviyesinde 14,22-kat olarak belirgin bir anlamlı artış olduğu belirlendi (p<0,01). Diğer yandan EMT sürecinde yer aldığı bilinen Snail, Twist ve Vimentin mRNA ifade düzeyinin artan RUT konsantrasyonuna bağlı olarak azaldığı ortaya konuldu (p<0,01).
Sonuç: RUT’un epitelden mezenkimal geçişi engelleme ve apoptozu teşvik edici potansiyelinin varlığı ortaya konulmuştur. Ayrıca RUT’un EMT üzerine olan etkilerinin daha ileri moleküler analizler ve in vivo olarak araştırılması önerilmektedir.
Anahtar Kelimeler
Kaynakça
- 1. Nouri Z, Fakhri S, Nouri K, Wallace CE, Farzaei MH, Bishayee A. Targeting Multiple Signaling Pathways in Cancer: The Rutin Therapeutic Approach. Cancers (Basel). 2020;12(8):2276. doi:10.3390/cancers12082276
- 2. Salehi B, Fokou PVT, Yamthe LRT, et al. Phytochemicals in Prostate Cancer: From Bioactive Molecules to Upcoming Therapeutic Agents. Nutrients. 2019;11(7):1483. doi:10.3390/nu11071483
- 3. Ganeshpurkar A, Saluja AK. The Pharmacological Potential of Rutin. Saudi Pharm J. 2017;25(2):149-164. doi:10.1016/j.jsps.2016.04.025
- 4. Li X, Chen G, Zhang X, et al. A new class of flavonol-based anti-prostate cancer agents: Design, synthesis, and evaluation in cell models. Bioorg Med Chem Lett. 2016;26(17):4241-4245. doi:10.1016/j.bmcl.2016.07.050
- 5. Pivec T, Kargl R, Maver U, et al. Chemical Structure-Antioxidant Activity Relationship of Water-Based Enzymatic Polymerized Rutin and Its Wound Healing Potential. Polymers (Basel). 2019;11(10):1566. doi:10.3390/polym11101566
- 6. Satari A, Ghasemi S, Habtemariam S, Asgharian S, Lorigooini Z. Rutin: A Flavonoid as an Effective Sensitizer for Anticancer Therapy; Insights into Multifaceted Mechanisms and Applicability for Combination Therapy. Evid Based Complement Alternat Med. 2021;2021:9913179. Published 2021 Aug 23. doi:10.1155/2021/9913179
- 7. Farha AK, Gan RY, Li HB, et al. The anticancer potential of the dietary polyphenol rutin: Current status, challenges, and perspectives. Crit Rev Food Sci Nutr. 2022;62(3):832-859. doi:10.1080/10408398.2020.1829541
- 8. Nieszporek A, Skrzypek K, Adamek G, Majka M. Molecular mechanisms of epithelial to mesenchymal transition in tumor metastasis. Acta Biochim Pol. 2019;66(4):509-520. doi:10.18388/abp.2019_2899
- 9. Odero-Marah V, Hawsawi O, Henderson V, Sweeney J. Epithelial-Mesenchymal Transition (EMT) and Prostate Cancer. Adv Exp Med Biol. 2018;1095:101-110. doi:10.1007/978-3-319-95693-0_6
- 10. Shibue T, Weinberg RA. EMT, CSCs, and drug resistance: the mechanistic link and clinical implications. Nat Rev Clin Oncol. 2017;14(10):611-629. doi:10.1038/nrclinonc.2017.44
- 11. Lu W, Kang Y. Epithelial-Mesenchymal Plasticity in Cancer Progression and Metastasis. Dev Cell. 2019;49(3):361-374. doi:10.1016/j.devcel.2019.04.010
- 12. Talebi H, Farahpour MR, Hamishehkar H. The effectiveness of Rutin for prevention of surgical induced endometriosis development in a rat model. Sci Rep. 2021;11(1):7180.. doi:10.1038/s41598-021-86586-4
- 13. Subhawa S, Naiki-Ito A, Kato H, et al. Suppressive Effect and Molecular Mechanism of Houttuynia cordata Thunb. Extract against Prostate Carcinogenesis and Castration-Resistant Prostate Cancer. Cancers (Basel). 2021;13(14):3403. doi:10.3390/cancers13143403
DETERMINATION OF THE EFFECT OF RUTIN ON EPITHHELIAL AND MESENCHIMAL TRANSITION IN PROSTATE CANCER CELLS
Öz
Objective: The difficulties experienced in the treatment of prostate cancer and the excess of side effects due to chemotherapy have brought the search for alternative treatment strategies. In recent studies, it is known that Rutin (RUT) has an anti-cancer effect on cancer cells. Our study aimed to determine the effects of RUT on epithelial-mesenchymal transition (EMT) in prostate cancer cells, for the first time.
Methods: The anticancer activity of RUT in prostate cancer cells (PC-3) was determined by WST-1, Annexin V ELISA, DAPI and Acridine Orange staining, and the anti-cancer and anti-metastatic properties of RUT were evaluated with the Scratch Assay test. The mRNA expression level of Bax, Bcl-2, Snail, Twist, Vimentin and E-cadherin genes was determined by RT-PCR.
Results: PC-3 cells were treated with RUT (500, 750, 1000, 1500 µM) for 24 and 48 hours. The viability rates decreased with increasing RUT concentration depending on dose and time (p<0.01). Annexin V protein decreased, and apoptotic cells increased depending on the increasing RUT concentration (p<0.01). There was a decrease in the number of migrating cells, especially at 750 and 1000 µM RUT concentrations. There was a significant increase (14.22-fold) in E-cadherin mRNA level after 1500 µM RUT treatment (p<0.01). On the other hand, the mRNA expression level of Snail, Twist and Vimentin, decreased at higher RUT concentrations (p<0.01).
Conclusion: The existence of RUT's potential to inhibit epithelial-mesenchymal transition and promote apoptosis has been demonstrated. It is also recommended to investigate the effects of RUT on EMT in vivo.
Anahtar Kelimeler
Prostate cancer rutin epithelial mesenchymal transition apoptosis
Kaynakça
- 1. Nouri Z, Fakhri S, Nouri K, Wallace CE, Farzaei MH, Bishayee A. Targeting Multiple Signaling Pathways in Cancer: The Rutin Therapeutic Approach. Cancers (Basel). 2020;12(8):2276. doi:10.3390/cancers12082276
- 2. Salehi B, Fokou PVT, Yamthe LRT, et al. Phytochemicals in Prostate Cancer: From Bioactive Molecules to Upcoming Therapeutic Agents. Nutrients. 2019;11(7):1483. doi:10.3390/nu11071483
- 3. Ganeshpurkar A, Saluja AK. The Pharmacological Potential of Rutin. Saudi Pharm J. 2017;25(2):149-164. doi:10.1016/j.jsps.2016.04.025
- 4. Li X, Chen G, Zhang X, et al. A new class of flavonol-based anti-prostate cancer agents: Design, synthesis, and evaluation in cell models. Bioorg Med Chem Lett. 2016;26(17):4241-4245. doi:10.1016/j.bmcl.2016.07.050
- 5. Pivec T, Kargl R, Maver U, et al. Chemical Structure-Antioxidant Activity Relationship of Water-Based Enzymatic Polymerized Rutin and Its Wound Healing Potential. Polymers (Basel). 2019;11(10):1566. doi:10.3390/polym11101566
- 6. Satari A, Ghasemi S, Habtemariam S, Asgharian S, Lorigooini Z. Rutin: A Flavonoid as an Effective Sensitizer for Anticancer Therapy; Insights into Multifaceted Mechanisms and Applicability for Combination Therapy. Evid Based Complement Alternat Med. 2021;2021:9913179. Published 2021 Aug 23. doi:10.1155/2021/9913179
- 7. Farha AK, Gan RY, Li HB, et al. The anticancer potential of the dietary polyphenol rutin: Current status, challenges, and perspectives. Crit Rev Food Sci Nutr. 2022;62(3):832-859. doi:10.1080/10408398.2020.1829541
- 8. Nieszporek A, Skrzypek K, Adamek G, Majka M. Molecular mechanisms of epithelial to mesenchymal transition in tumor metastasis. Acta Biochim Pol. 2019;66(4):509-520. doi:10.18388/abp.2019_2899
- 9. Odero-Marah V, Hawsawi O, Henderson V, Sweeney J. Epithelial-Mesenchymal Transition (EMT) and Prostate Cancer. Adv Exp Med Biol. 2018;1095:101-110. doi:10.1007/978-3-319-95693-0_6
- 10. Shibue T, Weinberg RA. EMT, CSCs, and drug resistance: the mechanistic link and clinical implications. Nat Rev Clin Oncol. 2017;14(10):611-629. doi:10.1038/nrclinonc.2017.44
- 11. Lu W, Kang Y. Epithelial-Mesenchymal Plasticity in Cancer Progression and Metastasis. Dev Cell. 2019;49(3):361-374. doi:10.1016/j.devcel.2019.04.010
- 12. Talebi H, Farahpour MR, Hamishehkar H. The effectiveness of Rutin for prevention of surgical induced endometriosis development in a rat model. Sci Rep. 2021;11(1):7180.. doi:10.1038/s41598-021-86586-4
- 13. Subhawa S, Naiki-Ito A, Kato H, et al. Suppressive Effect and Molecular Mechanism of Houttuynia cordata Thunb. Extract against Prostate Carcinogenesis and Castration-Resistant Prostate Cancer. Cancers (Basel). 2021;13(14):3403. doi:10.3390/cancers13143403
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Biyokimya ve Hücre Biyolojisi (Diğer) |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 28 Şubat 2023 |
| Gönderilme Tarihi | 6 Eylül 2022 |
| Kabul Tarihi | 1 Aralık 2022 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 6 Sayı: 1 |
"Acta Medica Nicomedia" Tıp dergisinde https://dergipark.org.tr/tr/pub/actamednicomedia adresinden yayımlanan makaleler açık erişime sahip olup Creative Commons Atıf-AynıLisanslaPaylaş 4.0 Uluslararası Lisansı (CC BY SA 4.0) ile lisanslanmıştır.