ÜÇLÜ NEGATİF MEME KANSERİNDE (TNBC), AZD7762 İLACINA KARŞI HASSASLIĞIN RBFOX2 GEN EKSPRESYONU İLE ARAŞTIRILMASI

Year 2022, , 33 - 37, 22.08.2022

İsmail Mert Alkaç , Murat İşbilen , Barış Küçükkaraduman , Ali Osmay Güre , Burçak Vural

https://doi.org/10.26650/experimed.1082020

Abstract

Amaç
Üçlü negative meme kanseri (ÜNMK), meme kanserleri arasında en agresif, metastatik ve prognozu kötü takip eden gruptur. ÜNMK’ne karşı moleküler hedefe yönelik standart bir tedavi yaklaşımı henüz mevcut değildir. Bu nedenle yeni terapötikler geliştirilmelidir. Bu çalışmada, AZD7762'nin birkaç meme kanseri hücre hattı üzerindeki etkisini belirlemeyi ve bu ilaca duyarlılığı göstermek için bir belirteç olarak RBFOX2 gen ekspresyon düzeylerini değerlendirmeyi amaçladık.
Materyal ve metod
AZD7762'nin meme kanseri hücre dizileri üzerindeki sitotoksik etkisi Sulforhodamine B yöntemi ile belirlendi. RBFOX2 geninin ekspresyon seviyeleri qPCR ile belirlendi. Seçilen ilaç AZD7762'nin IC50 değerleri ile RBFOX2 ekspresyonu arasındaki korelasyonu değerlendirmek için Pearson r korelasyon analizi yapıldı.
Bulgular
ÜNMK ve ÜNMK olmayan grupların IC50 değerleri arasında anlamlı bir fark olmamasına rağmen, ÜNMK hücre hatlarının AZD7762'ye daha duyarlı olma eğiliminde olduğu belirlendi. Ek olarak, AZD7762'ye daha fazla duyarlılık gösteren hücrelerde artan RBFOX2 ekspresyon seviyelerinin tespit edildi.
Sonuçlar
RBFOX2 geninin AZD7762 etkinliğini göstermek için bir biyobelirteç olarak kullanılabileceği sonucuna varılmıştır. AZD7762'nin etkisiyle ilişkili potansiyel sinyal mekanizmalarını araştırmak için daha fazla çalışmaya ihtiyaç vardı.

Keywords

Üçlü negatif meme kanseri, RBFOX2, AZD7762, Biyobelirteç

Investigation of Sensitivity to AZD7762 in Triple-Negative Breast Cancer (TNBC) with RBFOX2 Gene Expression as a Biomarker

Abstract

Objective: Triple negative breast cancer (TNBC) is one of the most metastatic, aggressive with poor prognosis types of breast cancers. There is currently no standard molecular-targeted treatment for TNBC. Therefore, new therapeutics should be developed. The aim of this study was to determine the effect of AZD7762 on several breast cancer cell lines and evaluate the RBFOX2 gene expression levels as a marker to show sensitivity to this drug.Materials and Methods: The cytotoxic effect of AZD7762 on breast cancer cell lines was determined by sulforhodamine B method. The expression levels of RBFOX2 gene were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The association between the IC50 values of the selected drug AZD7762 and RBFOX2 expression levels was evaluated by using Pearson r correlation analysis.Results: Although there was no significant difference between the IC50 values of TNBC and non-TNBC groups, it was determined that TNBC cell lines tended to be more sensitive to AZD7762. In addition, it was obvious that increasing levels of RBFOX2 expression were detected in cells that showed more sensitivity to AZD7762. Conclusion: It was concluded that the RBFOX2 gene can be used as a biomarker to show AZD7762 efficiency. Further studies are needed to investigate the potential signaling mechanisms that are associated with the effect of AZD7762.

Keywords

Triple Negative Breast Cancer, AZD7762, RBFOX2, Biomarker

References

• 1. Ferlay J, Colombet M, Soerjomataram I, Parkin DM, Pineros M, Znaor A, et al. Cancer statistics for the year 2020: An overview. Int J Cancer 2021; 149(4): 778-89. google scholar
• 2. Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2021. CA Cancer J Clin.2022; 72(1): 7-33. [CrossRef] google scholar
• 3. World Health Organization, Globocan 2020 database, Turkey can-cer statistics. (Available from: https://gco.iarc.fr/today/data/fact-sheets/populations/792-turkey-fact-sheets.pdf) google scholar
• 4. Karagoz Eren S, Arslan A, Akay E, Özhan N, Dönder Y. Assessing Clinicopathological features and prognosis of triple-negative breast cancer patients: A single-center study in Turkey. Arch Breast Cancer 2021; 8(4): 297-304. [CrossRef] google scholar
• 5. Landry I, Sumbly V, Vest M. Advancements in the treatment of triple-negative breast cancer: A narrative review of the literature. Cureus 2022; 14(2). google scholar
• 6. Odemis DA, Celik B, Erciyas SK, Erdogan OS, Tuncer SB, Gultaslar BK, et al. Evaluation of BRCA1/2 gene mutations in patients with high-risk breast and/or ovarian cancer in Turkey. Turk J Biochem 2022. [CrossRef] google scholar
• 7. Wahba HA, El-Hadaad HA. Current approaches in treatment of tri-ple-negative breast cancer. Cancer Biol Med 2015; 12(2): 106-16. google scholar
• 8. Arslan C, Dizdar O, Altundag K. Pharmacotherapy of triple-nega-tive breast cancer. Expert Opin Pharmacother 2009; 10(13): 208193. [CrossRef] google scholar
• 9. Ma CX, Cai S, Li S, Ryan CE, Guo Z, Schaiff WT, et al. Targeting Chk1 in p53-deficient triple-negative breast cancer is therapeutically beneficial in human-in-mouse tumor models. J Clin Invest 2012; 122(4): 1541-52. [CrossRef] google scholar
• 10. Isono M, Hoffmann MJ, Pinkerneil M, Sato A, Michaelis M, Cinatl J, et al. Checkpoint kinase inhibitor AZD7762 strongly sensitises urothelial carcinoma cells to gemcitabine. J Exp Clin Cancer Res 2017; 36(1): 1-12. [CrossRef] google scholar
• 11. Sausville E, LoRusso P, Carducci M, Carter J, Quinn MF, Malburg L, et al. Phase I dose-escalation study of AZD7762, a checkpoint kinase inhibitor, in combination with gemcitabine in US patients with advanced solid tumors. Cancer Chemother Pharmacol 2014; 73(3): 539-49. [CrossRef] google scholar
• 12. Bryant C, Rawlinson R, Massey AJ. Chk1 Inhibition as a novel ther-apeutic strategy for treating triple-negative breast and ovarian cancers. BMC Cancer 2014; 14(1): 1-14. [CrossRef] google scholar
• 13. Zhu J, Zou H, Yu W, Huang Y, Liu B, Li T, et al. Checkpoint kinase inhibitor AZD7762 enhance cisplatin-induced apoptosis in osteo-sarcoma cells. Cancer Cell Int 2019; 19(1): 1-11. [CrossRef] google scholar
• 14. Zhang Y, Lai J, Du Z, Gao J, Yang S, Gorityala S, et al. Targeting ra-dioresistant breast cancer cells by single agent CHK1 inhibitor via enhancing replication stress. Oncotarget 2016; 7(23): 34688-702. [CrossRef] google scholar
• 15. Min DJ, He S, Green JE. Birinapant (TL32711) improves responses to GEM/AZD7762 combination therapy in triple-negative breast cancer cell lines. Anticancer Res 2016; 36(6): 2649-57. google scholar
• 16. Garnett MJ, Edelman EJ, Heidorn SJ, Greenman CD, Dastur A, Lau KW, et al. Systematic identification of genomic markers of drug sensitivity in cancer cells. Nature 2012; 483(7391): 570-5. [Cross-Ref] google scholar
• 17. Kao J, Salari K, Bocanegra M, Choi Y La, Girard L, Gandhi J, et al. Mo-lecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PLoS ONE 2009; 4(7): e6146. google scholar
• 18. Liu X, Sun C, Jin X, Li P, Ye F, Zhao T, et al. Genistein enhances the radiosensitivity of breast cancer cells via G2/M cell cycle arrest and apoptosis. Molecules 2013;18(11): 13200-17. [CrossRef] google scholar
• 19. Ma Z, Yao G, Zhou B, Fan Y, Gao S, Feng X. The Chk1 inhibitor AZD7762 sensitises p53 mutant breast cancer cells to radiation in vitro and in vivo. Mol Med Rep 2012; 6(4): 897-903. [CrossRef] google scholar
• 20. Zhu J, Zou H, Yu W, Huang Y, Liu B, Li T, et al. Checkpoint kinase inhibitor AZD7762 enhance cisplatin-induced apoptosis in osteo-sarcoma cells. Cancer Cell Int 2019; 19(1): 195. google scholar
• 21. Liu Y, Li Y, Wang X, Liu F, Gao P, Quinn MM, et al. Gemcitabine and Chk1 inhibitor AZD7762 synergistically suppress the growth of Lkb1-deficient lung adenocarcinoma. Cancer Res 2017; 77(18): 5068-76. [CrossRef] google scholar
• 22. Bennett CN, Tomlinson CC, Michalowski AM, Chu IM, Luger D, Mittereder LR, et al. Cross-species genomic and functional anal-yses identify a combination therapy using a CHK1 inhibitor and a ribonucleotide reductase inhibitor to treat triple-negative breast cancer. Breast Cancer Res 2012; 14(4): R109. google scholar
• 23. Park JS, Lee CH, Kim HK, Kim D, Son JB, Ko E, et al. Suppression of the metastatic spread of breast cancer by DN10764 (AZD7762)-medi-ated inhibition of AXL signaling. Oncotarget 2016; 7(50): 8330818. [CrossRef] google scholar
• 24. Zhang Y, Lai J, Du Z, Gao J, Yang S, Gorityala S, et al. Targeting ra-dioresistant breast cancer cells by single agent CHK1 inhibitor via enhancing replication stress. Oncotarget 2016; 7(23): 34688-702. google scholar
• 25. Fici P, Gallerani G, Morel AP, Mercatali L, Ibrahim T, Scarpi E, et al. Splicing factor ratio as an index of epithelial-mesenchymal transi-tion and tumor aggressiveness in breast cancer. Oncotarget 2017; 8(2): 2423-36. [CrossRef] google scholar
• 26. Meng X, Yang S, Zhang J, Yu H. Contribution of alternative splicing to breast cancer metastasis. J Cancer Metastasis Treat 2019; 5: 21. [CrossRef] google scholar
• 27. Braeutigam C, Rago L, Rolke A, Waldmeier L, Christofori G, Win-ter J. The RNA-binding protein Rbfox2: An essential regulator of EMT-driven alternative splicing and a mediator of cellular inva-sion. Oncogene 2014; 33(9): 1082-92. [CrossRef] google scholar
• 28. Cen YL, Qi ML, Li HG, Su Y, Chen LJ, Lin Y, et al. Associations of polymorphisms in the genes of FGFR2, FGF1, and RBFOX2 with breast cancer risk by estrogen/progesterone receptor status. Mol Carcinog 2013; 52(SUPPL1): E52-9. google scholar
• 29. Oza V, Ashwell S, Almeida L, Brassil P, Breed J, Deng C, et al. Dis-covery of checkpoint kinase inhibitor (S)-5-(3-fluorophenyl)- N -(piperidin-3-yl)-3-ureidothiophene-2-carboxamide (AZD7762) by structure-based design and optimization of thiophenecarbox-amide ureas. J Med Chem 2012; 55(11): 5130-42. google scholar
• 30. Mitchell JB, Choudhuri R, Fabre K, Sowers AL, Citrin D, Zabludoff SD, et al. In vitro and in vivo radiation sensitization of human tu-mor cells by a novel checkpoint kinase inhibitor, AZD7762. Clin Cancer Res 2010; 16(7): 2076-84. [CrossRef] google scholar