Dosetaksel Dirençli Prostat Kanseri Hücrelerinde Timokinon Tarafından Otofajik Hücre Ölümünün İndüklenmesi

Year 2021, Volume: 23 Issue: 2, 187 - 191, 30.08.2021

Süleyman İlhan , Ferdi Oguz

https://doi.org/10.18678/dtfd.925238

Cited By: 3

Abstract

Amaç: Prostat kanserinde (prostate cancer, PCa) edinilen dosetaksel (docetaxel, DOC) direnci hala klinik bir sorundur. Kemoterapi tedavisinde başarısızlığa ek olarak tümör nüksüne neden olmaktadır. Bu nedenle, DOC’a dirençli PCa tedavisinde yeni ve daha etkili bileşiklere ihtiyaç duyulmaktadır. Bu çalışmanın amacı, Nigella sativa L. bitkisinin etken bileşenlerinden biri olan timokinon (thymoquinone, TQ)’un, DOC dirençli prostat kanseri hücreleri üzerindeki olası sitotoksik ve hücre ölümünü tetikleyici aktivitelerinin araştırılmasıdır.
Gereç ve Yöntemler: DOC dirençli PC3 hücreleri (DOC-R/PC3), büyüme ve bölünme yeteneklerini geliştirene kadar artan DOC (1-10 nM) konsantrasyonlarında devamlı kültürle çoğaltıldı. Hücre canlılığı, MTT yöntemi kullanılarak belirlendi. MuseTM Annexin V & Dead Cell kiti, apoptotik hücre ölümünün tespiti için kullanıldı. Otofajik vakuoller spesifik boya kullanılarak gösterildi. TQ mualemesi sonucu LC3-I, LC3-II ve Beclin-1 protein düzeylerindeki değişiklikler western blot analizi ile araştırıldı.
Bulgular: TQ muamelesi, DOC-R/PC3 hücrelerinin canlılığını doza ve zamana bağlı olarak inhibe etti (p=0.014). DOC-R/PC3 için TQ'nun IC50 değeri 72. saatte 60 µM olarak hesaplandı. TQ uygulaması, DOC dirençli prostat kanseri hücrelerinde apoptotik hücre ölümünü indüklemedi, ancak otofajik vakuol oluşumunu indükledi. Ayrıca TQ ile muamele edilen DOC-R/PC3 hücrelerinde Beclin-1 ve LC3-II protein seviyelerinin arttığı, ancak DOC-R/PC3 hücrelerinde LC3-I seviyelerinin azaldığı tespit edildi.
Sonuç: Tüm bu sonuçlar, TQ'nun gelecekte DOC dirençli prostat kanseri için yeni bir terapötik ajan olabileceğini göstermektedir.

Keywords

otofaji, Beclin-1, LC3, prostat kanseri, direnç, timokinon

Induction of Autophagic Cell Death by Thymoquinone in Docetaxel Resistant Prostate Cancer Cells

Abstract

Aim: Acquired docetaxel (DOC) resistance of prostate cancer (PCa) is still a clinical problem. In addition to failure in chemotherapy treatment, it causes tumor recurrence. Therefore, novel and more effective compounds are needed in DOC-resistant PCa treatment. This study aimed to investigate the possible cytotoxic and cell death-inducing activities of thymoquinone (TQ), one of the main active components of Nigella sativa L., on DOC-resistant prostate cancer cells.
Material and Methods: DOC-resistant PC3 cells (DOC-R/PC3) were developed by the continuous culture with increment concentrations of DOC (1-10 nM) until they improved their growth and division abilities. The cell viability was determined by MTT assay. The MuseTM Annexin V & Dead Cell kit was performed to detect apoptotic cell death. Autophagic vacuoles were observed by staining autophagic vacuoles. The levels of LC3I, LC3II and Beclin-1 proteins were investigated via western blot analysis.
Results: TQ inhibited the viability of DOC-R/PC3 cells in a dose- and time-dependent manner (p=0.014). The IC50 value of TQ for DOC-R/PC3 cells was calculated as 60 µM at 72 h. Treatment of TQ did not induce apoptotic cell death in DOC-resistant prostate cancer cells but induced the formation of autophagic vacuoles. Moreover, Beclin-1 and LC3-II protein levels were increased in TQ-treated DOC-R/PC3 cells, however, LC3-I levels were decreased in DOC-R/PC3 cells.
Conclusion: All these results show that TQ may become a new therapeutic target for DOC-resistant prostate cancer in the future.

Keywords

autophagy, Beclin-1, LC3, prostate cancer, resistance, thymoquinone

References

• Barsouk A, Padala SA, Vakiti A, Mohammed A, Saginala K, Thandra KC, et al. Epidemiology, staging and management of prostate cancer. Med Sci (Basel). 2020;8(3):28.
• Sartor O, de Bono JS. Metastatic prostate cancer. N Engl J Med. 2018;378(7):645-57.
• Nevedomskaya E, Baumgart SJ, Haendler B. Recent advances in prostate cancer treatment and drug discovery. Int J Mol Sci. 2018;19(5):1359.
• Cánovas V, Puñal Y, Maggio V, Redondo E, Marín M, Mellado B, et al. Prostate tumor overexpressed-1 (PTOV1) promotes docetaxel-resistance and survival of castration resistant prostate cancer cells. Oncotarget. 2017;8(35):59165-80.
• Singh SK, Apata T, Gordetsky JB, Singh R. Docetaxel combined with thymoquinone induces apoptosis in prostate cancer cells via inhibition of the PI3K/AKT signaling pathway. Cancers (Basel). 2019;11(9):1390.
• Salomi MJ, Nair SC, Panikkar KR. Inhibitory effects of Nigella sativa and saffron (Crocus sativus) on chemical carcinogenesis in mice. Nutr Cancer. 1991;16(1):67-72.
• Mostofa AGM, Hossain MK, Basak D, Bin Sayeed MS. Thymoquinone as a potential adjuvant therapy for cancer treatment: Evidence from preclinical studies. Front Pharmacol. 2017;8:295.
• Peng L, Liu A, Shen Y, Xu HZ, Yang SZ, Ying XZ, et al. Antitumor and anti-angiogenesis effects of thymoquinone on osteosarcoma through the NF-κB pathway. Oncol Rep. 2013;29(2):571-8.
• Khan MA, Tania M, Wei C, Mei Z, Fu S, Cheng J, et al. Thymoquinone inhibits cancer metastasis by downregulating TWIST1 expression to reduce epithelial to mesenchymal transition. Oncotarget. 2015;6(23):19580-91.
• Imran M, Rauf A, Khan IA, Shahbaz M, Qaisrani TB, Fatmawati S, et al. Thymoquinone: A novel strategy to combat cancer: A review. Biomed Pharmacother. 2018;106:390-402.
• Velho‑Pereira R, Kumar A, Pandey BN, Jagtap AG, Mishra KP. Radiosensitization in human breast carcinoma cells by thymoquinone: role of cell cycle and apoptosis. Cell Biol Int. 2011;35(10):1025-9.
• Zhang L, Bai Y, Yang Y. Thymoquinone chemosensitizes colon cancer cells through inhibition of NF-κB. Oncol Lett. 2016;12(4):2840-5.
• Hanahan D, Weinberg RA. Hallmarks of cancer: The next generation. 2011;144(5):646-74.
• Chen S, Rehman SK, Zhang W, Wen A, Yao L, Zhang J. Autophagy is a therapeutic target in anticancer drug resistance. Biochim Biophys Acta. 2010;1806(2):220-9.
• Goldsmith J, Levine B, Debnath J. Autophagy and cancer metabolism. Methods Enzymol. 2014;542:25-57.
• Farrow JM, Yang JC, Evans CP. Autophagy as a modulator and target in prostate cancer. Nat Rev Urol. 2014;11(9):508-16.
• Atmaca H, İlhan S, Batır MB, Pulat ÇÇ, Güner A, Bektaş H. Novel benzimidazole derivatives: Synthesis, in vitro cytotoxicity, apoptosis and cell cycle studies. Chem Biol Interact. 2020;327:109163.
• Atmaca H, Camli Pulat C, Cittan M. Liquidambar orientalis Mill. gum extract induces autophagy via PI3K/Akt/mTOR signaling pathway in prostate cancer cells. Int J Environ Health Res. 2020;[Epub ahead of print]. doi: 10.1080/09603123.2020.1818187.
• Dirican A, Atmaca H, Bozkurt E, Erten C, Karaca B, Uslu R. Novel combination of docetaxel and thymoquinone induces synergistic cytotoxicity and apoptosis in DU-145 human prostate cancer cells by modulating PI3K-AKT pathway. Clin Transl Oncol. 2015;17(2):145-51.
• Dirican A, Erten C, Atmaca H, Bozkurt E, Kucukzeybek Y, Varol U, et al. Enhanced cytotoxicity and apoptosis by thymoquinone in combination with zoledronic acid in hormone- and drug-resistant prostate cancer cell lines. J BUON. 2014;19(4):1055-61.
• El-Mahdy MA, Zhu Q, Wang QE, Wani G, Wani AA. Thymoquinone induces apoptosis through activation of caspase-8 and mitochondrial events in p53-null myeloblastic leukemia HL-60 cells. Int J Cancer. 2005;117(3):409-17.
• Arafa el-SA, Zhu Q, Shah ZI, Wani G, Barakat BM, Racoma I, et al. Thymoquinone up-regulates PTEN expression and induces apoptosis in doxorubicin-resistant human breast cancer cells. Mutat Res. 2011;706(1-2):28-35.
• Taha MME, Sheikh BY, Salim LZA, Mohan S, Khan A, Kamalidehghan B, et al. Thymoquinone induces apoptosis and increase ROS in ovarian cancer cell line. Cell Mol Biol (Noisy-le-grand). 2016;62(6):97-101.
• Park JE, Kim DH, Ha E, Choi SM, Choi JS, Chun KS, et al. Thymoquinone induces apoptosis of human epidermoid carcinoma A431 cells through ROS-mediated suppression of STAT3. Chem Biol Interact. 2019;312:108799.
• Clarke PG. Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl). 1990;181(3):195-213.
• Wei Y, Pattingre S, Sinha S, Bassik M, Levine B. JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy. Mol Cell. 2008;30(6):678-88.
• Zalckvar E, Berissi H, Mizrachy L, Idelchuk Y, Koren I, Eisenstein M, et al. DAP-kinase-mediated phosphorylation on the BH3 domain of beclin 1 promotes dissociation of beclin 1 from Bcl-XL and induction of autophagy. EMBO Rep. 2009;10(3):285-92.
• Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, et al. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol. 2001;152(4):657-68.